TWI453209B - Compounds - Google Patents

Description

Compound

The present invention relates to a spirocyclic guanamine derivative, a process for the preparation thereof, a pharmaceutical composition containing the same, a process for preparing the same, a therapeutic use thereof, and an intermediate for use in the preparation thereof.

The first line of treatment for various pulmonary conditions, including chronic obstructive pulmonary disease (COPD) and asthma, is via the use of bronchodilators. A muscarinic receptor antagonist (anticholinergic drug) is a bronchodilator that functions by reducing vagal cholinergic activity (the main reversible component of respiratory contraction in COPD). Beta-adrenergic receptor agonists are also bronchodilators due to the bronchoconstriction response of a functional mediated mediated mediator (including acetylcholine).

In addition to improving lung function, these agents also improve dyspnea (gasp), quality of life, exercise load and reduce the severity of the disease. There have been many clinical studies demonstrating that combination of anticholinergic drugs and β ₂ -acceptor agonists is more effective than any individual component (van Noord, JA, Aumann, JL., Janssens, E., Smeets, JJ, Verhaert, J., Disse, B., Mueller, A. & Cornelissen, PJG, 2005. "Comparison of tiotropium once daily, formoterol twice daily and both combined once daily in patients with COPD", Eur. Respir. J. , vol 26 , pp 214-222.). The single molecule (MABA) active in muscarinic and beta ₂ receptors provides an additional benefit to COPD patients over the efficacy and side effect profiles of either individual agent. Moreover, molecules with dual activity can also provide advantages over the ease of use and patient compliance of co-administration of a single therapeutic agent. The monotherapy is also advantageous because it is more balanced than the two individual compounds. If combined with another treatment, it also provides the possibility of a triple therapeutic agent.

According to a first aspect of the invention, we now provide a compound of formula I

Wherein Ar represents a β-adrenergic receptor binding group; L represents a linker comprising a linear or branched hydrocarbon group having up to 15 carbon atoms; wherein up to three carbon atoms in the chain are optionally up to three Substituted by four substituents independently selected from the group consisting of halogen, S(O) _0-2 R ⁵⁶ , NR ⁵⁷ R ⁵⁸ , S(O) ₂ NR ⁵⁹ R ⁶⁰ , C(O)NR ⁶¹ R ⁶² , C( O) OR ⁶³ , NR ⁶⁴ S(O) ₂ R ⁶⁵ , NR ⁶⁶ C(O)R ⁶⁷ , NR ⁶⁸ C(O)OR ⁶⁹ , NR ⁷⁰ C(O)NR ⁷¹ R ⁷² , OR ⁷³ , C _{1- 6} alkyl and C _3-6 cycloalkyl, and wherein C _1-6 alkyl and C _3-6 cycloalkyl are optionally substituted with up to three substituents independently selected from the group consisting of halogen, hydroxy and C _{1 -6} alkoxy; wherein up to five carbon atoms in the chain may be replaced by groups independently selected from: O, NR ⁴⁵ , S, S(O), S(O) ₂ , C(O)O, OC(O), NR ⁴⁶ C(O), C(O)NR ⁴⁷ , NR ⁴⁸ S(O) ₂ , S(O) ₂ NR ⁴⁹ , NR ⁵⁰ C(O)NR ⁵¹ , NR ⁵² S(O) ₂ NR ⁵³ , OC(O)NR ⁵⁴ , NR ⁵⁵ C(O)O, with the proviso that any of the groups in the chain are separated by at least 2 carbon atoms; and wherein there are up to six carbon atoms in the chain Can be formed with up to four independent choices

a portion of a monocyclic or bicyclic aliphatic, heteroaliphatic, aromatic or heteroaromatic ring of a hetero atom of N, O or S, the ring comprising up to 10 ring atoms, and wherein the ring is up to three independent depending on the situation Substituted by a substituent selected from the group consisting of halogen, S(O) _0-2 R ⁵⁶ , NR ⁵⁷ R ⁵⁸ , S(O) ₂ NR ⁵⁹ R ⁶⁰ , C(O)NR ⁶¹ R ⁶² , C(O)OR ⁶³ , NR ⁶⁴ S(O) ₂ R ⁶⁵ , NR ⁶⁶ C(O)R ⁶⁷ , NR ⁶⁸ C(O)OR ⁶⁹ , NR ⁷⁰ C(O)NR ⁷¹ R ⁷² , OR ⁷³ , C _1-6 alkyl And a C _3-6 cycloalkyl group, wherein the C _1-6 alkyl group and the C _3-6 cycloalkyl group are optionally substituted with up to three substituents independently selected from the group consisting of halogen, hydroxy and C _1-6 alkane And the chain may comprise up to three independently selected such rings; wherein R ⁵⁶ , R ⁶⁵ and R ⁶⁹ each independently represent a C _1-6 alkyl group or a C _3-6 cycloalkyl group, wherein the C _{1 -6} alkyl and C _3-6 cycloalkyl may be optionally substituted with up to three substituents independently selected from halogen, hydroxy, C _1-6 alkoxy; and R ⁴⁵ , R ⁴⁶ , R ⁴⁷ , ^{R 48, R 49, R 50} , R 51, R 52, R 53, R 54, R 55, R 57, R 58, R 59, R 60, R 61, R 62, R 63, R 64, R 66 ^{R 67, R 68, R 70} , R 71, R 72 and R ⁷³ each independently represents hydrogen, C _1-6 alkyl or C _3-6 cycloalkyl wherein the C _1-6 alkyl and C _3-6 The cycloalkyl group may be optionally substituted with up to three substituents independently selected from halogen, hydroxy, _C1-6 alkoxy; or R ⁵⁷ and R ⁵⁸ , R ⁵⁹ and R ⁶⁰ , R ⁶¹ and R ⁶² or Any of R ⁷¹ and R ⁷² together with a nitrogen atom linking the two may form a 4 to 8 membered aliphatic heterocyclic ring, wherein the heterocyclic ring may comprise up to three heteroatoms independently selected from N, O and S, wherein The ring may optionally be substituted with up to three substituents independently selected from halogen, hydroxy, _C1-6 alkyl or _C3-6 cycloalkyl, wherein the _C1-6 alkyl and _C3-6 The cycloalkyl group may be optionally substituted with up to three substituents independently selected from the group consisting of halogen, hydroxy and C _1-6 alkoxy; and wherein the chain may additionally comprise up to three carbon-carbon double bonds; wherein the chain Up to three carbon-carbon ginsins may additionally be included; L ¹ and L ² independently represent hydrogen, C _1-6 alkyl or C _3-6 cycloalkyl; L ³ and L ⁴ independently represent hydrogen, C _{1- a 6} alkyl group or a C _3-6 cycloalkyl group, wherein the C _1-6 alkyl group and the C _3-6 cycloalkyl group In the case of up to three substituents independently selected from the group consisting of halogen, hydroxy and C _1-6 alkoxy; in addition, L ¹ and/or L ³ may be bonded to the carbon atom of the hydrocarbyl chain in the linker L to form An aliphatic ring having up to 6 ring atoms, wherein each ring may contain up to three heteroatoms independently selected from N, O and S; R ¹ represents a benzene ring, a 4 to 8 member heteroaliphatic ring, 3 to An 8-membered aliphatic ring or a 5 to 6 membered heteroaryl ring each having up to four heteroatoms independently selected from N, O or S, wherein in each case the ring may optionally be up to three independently selected from the group consisting of Substituted by a substituent: halogen, cyano, nitro, SH, S(O) _0-2 R ⁵ , NR ⁶ R ⁷ , S(O) ₂ NR ⁸ R ⁹ , C(O)NR ¹⁰ R ¹¹ , C (O)OR ¹² , NR ¹³ S(O) ₂ R ¹⁴ , NR ¹⁵ C(O)R ¹⁶ , NR ¹⁷ C(O)OR ¹⁸ , NR ¹⁹ C(O)NR ²⁰ R ²¹ , OR ²² , C _{1 -7} alkyl or C _3-8 cycloalkyl (in which case the C _1-7 alkyl and C _3-8 cycloalkyl may be optionally substituted by up to six substituents independently selected from: halogen , cyano, nitro, SH, S(O) _0-2 R ⁵ , NR ⁶ R ⁷ , S(O) ₂ NR ⁸ R ⁹ , C(O)NR ¹⁰ R ¹¹ , C(O)OR ¹² , NR ¹³ S(O) ₂ R ¹⁴ , NR ¹⁵ C(O)R ¹⁶ , NR ¹⁷ C(O)OR ¹⁸ , NR ¹⁹ C(O)NR ²⁰ R ²¹ , OR ²² ), benzene ring, 4 to 8 member heteroaliphatic ring, each having up to four a 5 to 6 membered heteroaryl ring independently selected from heteroatoms of N, O or S, a benzene ring, a 4 to 8 membered heteroaliphatic ring, a 3 to 8 membered aliphatic ring or a 5 to 6 membered heteroaryl ring Each of the cases may be substituted by up to three substituents independently selected from the group consisting of halogen, cyano, nitro, SH, S(O) _0-2 R ⁵ , S(O) ₂ NR ⁸ R ⁹ , C(O And NR ¹⁰ R ¹¹ , C(O)OR ¹² , OR ²² , C _1-6 alkyl or C _3-6 cycloalkyl, wherein the C _1-6 alkyl group and the C _3-6 cycloalkyl group may be used as the case may be Up to three substituents independently selected from the group consisting of halogen, hydroxy, C _1-6 alkoxy, cyano, nitro, NH ₂ , NH(C _1-6 alkyl) and N (C _1-6 Alkyl) ₂ ; or R ¹ represents a fused aliphatic, fused heteroaliphatic, fused aromatic or fused having up to 10 atoms and having up to four heteroatoms independently selected from N, O or S Heteroaryl ring, in each case wherein the ring may be substituted with up to three substituents independently selected from the group consisting of halogen, cyano, nitro, SH, S(O) _0-2 R ⁵ , NR ⁶ R ⁷ , S(O) ₂ NR ⁸ R ⁹ , C(O)NR ¹⁰ R ¹¹ , C(O)OR ¹² , NR ¹³ S(O) ₂ R ¹⁴ , NR ¹⁵ C(O)R ¹⁶ , NR ¹⁷ C(O)OR ¹⁸ , NR ¹⁹ C(O Or NR ²⁰ R ²¹ , OR ²² , C _1-6 alkyl or C _3-6 cycloalkyl, wherein the C _1-6 alkyl group and the C _3-6 cycloalkyl group may optionally be selected from up to three independently selected from Substituted by a substituent: halogen, hydroxy, C _1-6 alkoxy, NH ₂ , NH(C _1-6 alkyl) and N(C _1-6 alkyl) ₂ ; or R ¹ further represents C _{1- a 6} alkyl chain wherein one or two carbon atoms may be replaced by O, S or N and wherein R ¹ may be substituted with up to three C _1-3 alkyl chains and two of the chains may be joined to form C _{3 a -8} cycloalkyl chain, wherein the C _1-3 alkyl group and the C _3-8 cycloalkyl chain are optionally substituted with up to three substituents independently selected from the group consisting of halogen, cyano, nitro, SH, S(O) _0-2 R ⁵ , NR ⁶ R ⁷ , S(O) ₂ NR ⁸ R ⁹ , C(O)NR ¹⁰ R ¹¹ , C(O)OR ¹² , NR ¹³ S(O) ₂ R ¹⁴ NR ¹⁵ C(O)R ¹⁶ , NR ¹⁷ C(O)OR ¹⁸ , NR ¹⁹ C(O)NR ²⁰ R ²¹ , OR ²² , and wherein the C _1-6 alkyl chain further depends on the situation up to three Substituted independently by a substituent selected from the group consisting of halogen, cyano, nitro, SH, S(O) _0-2 R ⁵ NR ⁶ R ⁷ , S(O) ₂ NR ⁸ R ⁹ , C(O)NR ¹⁰ R ¹¹ , C(O)OR ¹² , NR ¹³ S(O) ₂ R ¹⁴ , NR ¹⁵ C(O)R ¹⁶ , NR ¹⁷ C(O)OR ¹⁸ , NR ¹⁹ C(O)NR ²⁰ R ²¹ , OR ²² , benzene ring, 4 to 8 member heteroaliphatic ring, 3 to 8 member aliphatic ring, each having up to four 5 to 6 membered heteroaryl rings independently selected from heteroatoms of N, O or S, and wherein each ring is optionally substituted with up to three substituents independently selected from the group consisting of halogen, cyano, nitro, S ( O) _0-2 R ⁵ , S(O) ₂ NR ⁸ R ⁹ , C(O)NR ¹⁰ R ¹¹ , C(O)OR ¹² , OR ²² , C _1-7 alkyl or C _3-7 naphthenic a base (in each case wherein the C _1-7 alkyl group and the C _3-7 cycloalkyl group are optionally substituted with up to six substituents independently selected from the group consisting of halogen, cyano, nitro, SH, S ( O) _0-2 R ⁵ , NR ⁶ R ⁷ , S(O) ₂ NR ⁸ R ⁹ , C(O)NR ¹⁰ R ¹¹ , C(O)OR ¹² , NR ¹³ S(O) ₂ R ¹⁴ , NR ¹⁵ C(O)R ¹⁶ , NR ¹⁷ C(O)OR ¹⁸ , NR ¹⁹ C(O)NR ²⁰ R ²¹ , OR ²² ), benzene ring, 4 to 8 member heteroaliphatic ring, 3 to 8 member aliphatic a 5 to 6 membered heteroaryl ring each having up to four heteroatoms independently selected from N, O or S, the benzene ring, a 4 to 8 member heteroaliphatic ring, and 3 to 8 member lipids Ring or a 5-6 heteroaryl each optionally ring by up to three independently selected from the substituents: halogen, cyano, ^{_{nitro, S (O) 0-2 R 5}} , S (O) 2 NR ⁸ R ⁹ , C(O)NR ¹⁰ R ¹¹ , C(O)OR ¹² , OR ²² , C _1-6 alkyl or C _3-6 cycloalkyl, wherein C _1-6 alkyl and C _3- Each of the ₆ cycloalkyl groups may be optionally substituted with up to three substituents independently selected from the group consisting of halogen, hydroxy, C _1-6 alkoxy, NH ₂ , NH(C _1-6 alkyl) and N (C _{1 -6} alkyl) ₂ ; or the C _1-6 alkyl chain may be fused, aliphatic or fused with up to 10 atoms and having up to four heteroatoms independently selected from N, O or S Substituted by an aliphatic, fused aromatic or fused heteroaryl ring, in each case wherein the ring may be substituted with up to three substituents independently selected from the group consisting of halogen, cyano, nitro, SH, S (O) _0-2 R ⁵ , NR ⁶ R ⁷ , S(O) ₂ NR ⁸ R ⁹ , C(O)NR ¹⁰ R ¹¹ , C(O)OR ¹² , NR ¹³ S(O) ₂ R ¹⁴ , NR ¹⁵ C(O)R ¹⁶ , NR ¹⁷ C(O)OR ¹⁸ , NR ¹⁹ C(O)NR ²⁰ R ²¹ , OR ²² , C _1-6 alkyl or C _3-6 cycloalkyl, wherein the C _1-6 alkyl and C _3-6 cycloalkyl may optionally be substituted with up to three independently From the group consisting of substituents: halogen, hydroxy, C _1-6 alkoxy, NH _2, NH (C _1-6 alkyl) and N (C _1-6 alkyl) _2; R ^5, R ^14, and R ¹⁸ independently represents a C _1-6 alkyl group or a C _3-6 cycloalkyl group, wherein the C _1-6 alkyl group and the C _3-6 cycloalkyl group may optionally be one or more substituents independently selected from the group consisting of Substituted: halogen, hydroxy, C _1-6 alkoxy, NH ₂ , NH(C _1-6 alkyl) and N(C _1-6 alkyl) ₂ ; and R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ , R ¹⁵ , R ¹⁶ , R ¹⁷ , R ¹⁹ , R ²⁰ , R ²¹ and R ²² each independently represent hydrogen, C _1-6 alkyl or C _3-6 a cycloalkyl group, wherein the C _1-6 alkyl group or the C _3-6 cycloalkyl group may be optionally substituted with one or more substituents independently selected from the group consisting of halogen, hydroxy, C _1-6 alkoxy, NH ₂ , NH(C _1-6 alkyl) and N(C _1-6 alkyl) ₂ ; or R ⁶ and R ⁷ , R ⁸ and R ⁹ , R ¹⁰ and R ¹¹ or R ²⁰ and R ²¹ The group may form a 4 to 8 membered aliphatic heterocyclic ring together with a nitrogen atom linking the two, wherein the heterocyclic ring may be optionally substituted with one or more substituents independently selected from the group consisting of halogen, hydroxy, C _1-6 alkane , C _1-6 cycloalkyl and C _1-6 alkoxy Wherein the C _1-6 alkyl group, the C _1-6 cycloalkyl group or the C _1-6 alkoxy group may be optionally substituted with one or more substituents independently selected from the group consisting of halogen, hydroxy and C _1-6 Alkoxy, NH ₂ , NH(C _1-6 alkyl) and N(C _1-6 alkyl) ₂ ; X represents O, S, S(O) _o or CR ²⁵ R ²⁶ ; m=0, 1 , 2 or 3; n = 1, 2, 3 or 4; the constraint is that m + n is greater than or equal to 2o = 1, 2; W represents CR ²⁷ R ²⁸ CR ²⁹ R ³⁰ or CR ³¹ R ³² CR ³³ R ³⁴ CR ³⁵ R ³⁶ ; V and Z independently represent the key, CR ³⁷ R ³⁸ or CR ³⁹ R ⁴⁰ CR ⁴¹ R ⁴² with the constraint that when X represents O, S or S(O) _o , m, V and The Z system is such that all heteroatoms in the ring are separated by at least two carbon atoms (for example, when V is a bond, m is not 0, Z is not a bond); Y represents CO, CONR ⁴³ , SO ₂ or SO ₂ NR ⁴⁴ ; R ²⁵ , R ²⁶ , R ²⁷ , R ²⁸ , R ²⁹ , R ³⁰ , R ³¹ , R ³² , R ³³ , R ³⁴ , R ³⁵ , R ³⁶ , R ³⁷ , R ³⁸ , R ³⁹ , R ⁴⁰ , R ⁴¹ , R ⁴² each independently represents hydrogen, fluorine or C _1-6 alkyl or C _3-6 cycloalkyl; and when it does not represent hydrogen or fluorine, R ²⁵ and R ²⁶ , R ²⁷ and R ²⁸ , R ²⁹ And R ³⁰ , R ³¹ and R ³² , R ³³ and R ³⁴ , R ³⁵ and R ³⁶ , R ³⁷ and R ³⁸ , R ³⁹ and R ⁴⁰ or R ⁴¹ and R ⁴² together with a carbon atom linking the two may additionally form a 3 to 6 membered aliphatic ring; R ⁴³ and R ⁴⁴ each independently represent hydrogen, C _1-6 alkane Or a C _3-6 cycloalkyl group and a pharmaceutically acceptable salt thereof.

The term "β-adrenergic receptor binding group" denotes a group which binds to a β-adrenergic receptor; such as, for example, the review article "β-adrenergic receptors in Comprehensive Medicinal Chemistry, 1990, BE Main, p187 (Pergamon Press) The groups are known from, for example, WO/2005092841, US/20050215542, WO/2005070872, WO/2006023460, WO/2006051373, WO/2006087315, WO/2006032627.

Suitable β-adrenergic receptor binding groups include

M ¹ is S, C(O), NA ⁵ , CA ⁶ A ⁷ , CH ₂ CH ₂ , CH=CH, CH ₂ O or OCH ₂ ; M ² is S, C(O), NA ⁵ , CA ⁶ A ⁷ , CH ₂ CH ₂ , CH=CH, CH ₂ O or OCH ₂ ; A ¹ , A ² , A ³ and A ^{4 are} independently hydrogen, halogen, trifluoromethyl, cyano, carboxyl, hydroxy, Nitro, S(O) ₂ A ⁸ , NA ⁹ S(O) ₂ A ¹⁰ , C(O)NA ¹¹ A ¹² , NA ¹³ C(O)A ¹⁴ , C _1-6 alkyl, C _1-6 Alkoxy, C(O)(C _1-6 alkyl) or C(O)OC _1-6 alkyl; A ³ may also be CH ₂ OH, NHCHO, NHS(O) ₂ NA ¹⁵ A ¹⁶ or NHSO ₂ A ¹⁷ ; A ⁵ , A ⁶ , A ⁷ , A ⁹ , A ¹¹ , A ¹² , A ¹³ , A ¹⁴ , A ¹⁵ or A ^{16 are} independently hydrogen or C _1-6 alkyl; A ⁸ , A ¹⁰ And A ^{17 is} independently a C _1-6 alkyl group; more preferably a β adrenergic receptor binding group Ar is selected from

Wherein M ¹ is S, CH=CH, CH ₂ O or OCH ₂ ; M ² is S, CH=CH, CH ₂ O or OCH ₂ ; A ¹ , A ² and A ^{4 are} independently hydrogen, halogen, C _1-6 alkyl, C _1-6 alkoxy; A ³ may also be CH ₂ OH, NHCHO, NHS(O) ₂ NA ¹⁵ A ¹⁶ or NHSO ₂ A ¹⁷ ; A ¹⁵ or A ¹⁶ is independently selected from Hydrogen, C _1-6 alkyl or C _3-6 cycloalkyl; A ¹⁷ is C _1-6 alkyl or C _3-6 cycloalkyl;

Examples of the C _1-6 alkyl group include a C _1-4 alkyl group and a C _1-2 alkyl group.

Examples of the C _3-6 cycloalkyl group include a C _3-5 cycloalkyl group and a C _3-4 cycloalkyl group.

Examples of the C _1-6 alkoxy group include a C _1-4 alkoxy group and a C _1-2 alkoxy group.

More suitable for the beta adrenergic receptor binding group Ar is selected from:

Wherein A ¹ , A ² and A ⁴ are all hydrogen, A ³ is CH ₂ OH, NHCHO, M ¹ is S, CH=CH or OCH ₂ ; M ² is S, CH=CH or OCH ₂ .

More suitable for the beta adrenergic receptor binding group Ar is selected from:

Suitably L represents a linker comprising up to 12 carbon atoms or up to 10 carbon atoms or up to 8 carbon atoms; wherein up to two carbon atoms in the chain are optionally independently selected from four up to four Substituted by the following substituents: halogen, S(O) _0-2 R ⁵⁶ , NR ⁵⁷ R ⁵⁸ , S(O) ₂ NR ⁵⁹ R ⁶⁰ , C(O)NR ⁶¹ R ⁶² , C(O)OR ⁶³ , NR ⁶⁴ S(O) ₂ R ⁶⁵ , NR ⁶⁶ C(O)R ⁶⁷ , NR ⁶⁸ C(O)OR ⁶⁹ , NR ⁷⁰ C(O)NR ⁷¹ R ⁷² , OR ⁷³ , C _1-6 alkyl and C _3-6 cycloalkyl, and wherein C _1-6 alkyl and C _3-6 cycloalkyl are optionally substituted with up to three substituents independently selected from the group consisting of halogen, hydroxy and C _1-6 alkoxy Suitably the up to two carbon atoms of the chain may be replaced by groups independently selected from: O, NR ⁴⁵ , S, S(O), S(O) ₂ , C(O)O, OC ( O), NR ⁴⁶ C(O), C(O)NR ⁴⁷ , NR ⁴⁸ S(O) ₂ , S(O) ₂ NR ⁴⁹ , NR ⁵⁰ C(O)NR ⁵¹ , NR ⁵² S(O) ₂ NR ⁵³ ; or independently selected from O, S, S(O), S(O) ₂ , NR ⁴⁶ C(O), C(O)NR ⁴⁷ ; the limitation is that in either case, any of the chains The group is separated by at least 2 carbon atoms; up to four carbon atoms in a suitable tether The moiety may form part of a monocyclic or bicyclic aliphatic, heteroaliphatic, aromatic or heteroaromatic ring having up to three heteroatoms independently selected from N, O or S, the ring comprising up to 10 ring atoms, and wherein the ring optionally by up to three substituents independently selected from the group substituted with: ^{_{halo, S (O) 0-2 R 56}} , NR 57 R 58, S (O) 2 NR 59 R 60, C (O) NR ⁶¹ R ⁶² , C(O)OR ⁶³ , NR ⁶⁴ S(O) ₂ R ⁶⁵ , NR ⁶⁶ C(O)R ⁶⁷ , NR ⁶⁸ C(O)OR ⁶⁹ , NR ⁷⁰ C(O)NR ⁷¹ R ⁷² , OR ⁷³ , C _1-6 alkyl and C _3-6 cycloalkyl, and wherein C _1-6 alkyl and C _3-6 cycloalkyl are optionally substituted by up to three substituents independently selected from Halogen, hydroxy and _C1-6 alkoxy; suitably the chain may comprise up to two or one independently selected such ring; suitably R ⁵⁶ , R ⁶⁵ and R ⁶⁹ each independently represent C _{1 a -4} alkyl or C _3-6 cycloalkyl group, wherein the C _1-4 alkyl group and the C _3-6 cycloalkyl group are optionally substituted with up to three substituents independently selected from the group consisting of halogen, hydroxyl, C _1-4 alkoxy; and suitably R ⁴⁵ , R ⁴⁶ , R ⁴⁷ , R ⁴⁸ , R ⁴⁹ , R ⁵⁰ , R ⁵¹ , R ⁵² , R ⁵³ , R ⁵⁴ , R ⁵⁵ , R ⁵⁷ , R ⁵⁸ , R ⁵⁹ , R ⁶⁰ , R ⁶¹ , R ⁶² , R ⁶³ , R ⁶⁴ , R ⁶⁶ , R ⁶⁷ , R ⁶⁸ , R ⁷⁰ , R ⁷¹ , R ⁷² and R ^{73 are} each independently represented Hydrogen, C _1-4 alkyl or C _3-6 cycloalkyl, wherein the C _1-4 alkyl and C _3-6 cycloalkyl are optionally substituted by up to three substituents independently selected from halogen: , a hydroxyl group, a C _1-4 alkoxy group; or any one of R ⁵⁷ and R ⁵⁸ , R ⁵⁹ and R ⁶⁰ , R ⁶¹ and R ⁶² or R ⁷¹ and R ⁷² together with a nitrogen atom connecting the two may form 4 To an 8-membered aliphatic heterocyclic ring, wherein the heterocyclic ring may be optionally substituted with up to three substituents independently selected from halogen, hydroxy and C _1-4 alkyl or C _3-6 cycloalkyl, wherein the C _1-4 alkyl and C _3-6 cycloalkyl may optionally be substituted with up to three independently selected from the substituents: halogen, hydroxy, and C _1-4 alkoxy; hydrocarbon group which may be present as part of the linker Examples of suitable ring systems include

Wherein the heterocyclyl ring is unsubstituted or substituted with 1 or 2 substituents independently selected from halogen: C _1-4 alkyl (substituted by the following groups: OR ¹²¹ , NR ¹²² R ¹²³ or NR ¹²⁴ C(O)R ¹²⁵ ), OR ¹²⁶ , NR ¹²⁷ R ¹²⁸ , C(O)NR ¹²⁹ R ¹³⁰ , NR ¹³¹ C(O)R ¹³² , CN, S(O) ₂ R ¹³³ or S( O) ₂ NR ¹³⁴ R ¹³⁵ ; R ¹³³ represents C _1-6 alkyl or C _3-6 cycloalkyl, wherein the C _1-6 alkyl group and the C _3-6 cycloalkyl group may optionally be independently selected by three from the group consisting of substituents: halogen, hydroxy, C _1-6 alkoxy group; and ^{R 121, R 122, R 123} , R 124, R 125, R 126, R 127, R 128, R 129, R 130 And R ¹³¹ , R ¹³² , R ¹³⁴ , R ¹³⁵ and R ¹³⁶ each independently represent hydrogen, C _1-6 alkyl or C _3-6 cycloalkyl, wherein the C _1-6 alkyl and C _3-6 cycloalkane The substituent may be substituted by up to three substituents independently selected from the group consisting of halogen, hydroxy, C _1-6 alkoxy; or R ¹²² and R ¹²³ , R ¹²⁷ and R ¹²⁸ , R ¹²⁹ and R ¹³⁰ or R ¹³⁴ R ¹³⁵ and to any group of two with the nitrogen atom, may form together 4-8 aliphatic heterocyclic ring, wherein the heterocyclic ring optionally substituted with up to three Independently selected from the substituents: halogen, hydroxy, C _1-6 alkyl or C _3-6 cycloalkyl wherein the C _1-6 alkyl and C _3-6 cycloalkyl optionally substituted with up to three Substituents independently selected from halogen and hydroxy are substituted; suitably the chain may additionally comprise up to two carbon-carbon double bonds or a single carbon-carbon double bond.

Suitably the chain may additionally comprise up to two carbon-carbon bonds or a single carbon-carbon bond.

Suitably L ¹ , L ² , L ³ and L ⁴ each independently represent hydrogen, C _1-4 alkyl or C _3-6 cycloalkyl, wherein the C _1-4 alkyl and C _3-6 cycloalkane The base may be substituted by up to three substituents independently selected from a halogen and a hydroxyl group; in addition, L ¹ and/or L ³ may be bonded to a carbon atom of the hydrocarbyl chain in the linker L to form up to 6 ring atoms. An aliphatic ring wherein the ring may contain up to two heteroatoms independently selected from N, O and S.

Wherein the four carbon atoms of the chain form part of a monocyclic or bicyclic aliphatic, heteroaliphatic, aromatic or heteroaromatic ring having up to three heteroatoms independently selected from N, O or S (if An aliphatic ring system) may contain up to 10, 9, 8, 7, 6, 5, 4 or 3 ring atoms; if it is an aromatic ring system, it may contain 10, 9, 6 or 5 ring atoms; Each line is independently selected; R ¹ represents a benzene ring or a 5 to 6 membered heteroaryl ring, each having up to three heteroatoms independently selected from N, O or S or R ¹ representing up to 10 atoms and having up to three a fused aliphatic, fused heteroaliphatic, fused aromatic or fused heteroaryl ring independently selected from heteroatoms of N, O or S, wherein each ring may optionally be up to three independently selected from the group consisting of Substituted by a substituent: halogen, cyano, nitro, SH, S(O) _0-2 R ⁵ , NR ⁶ R ⁷ , S(O) ₂ NR ⁸ R ⁹ , C(O)NR ¹⁰ R ¹¹ , C ^{(O) OR 12, NR 13} S (O) 2 R 14, NR 15 C (O) R 16, NR 17 C (O) OR 18, NR 19 C (O) NR 20 R 21, OR 22, C 1 _-7 alkyl group or a C _3-8 cycloalkyl group (wherein in each case the C _1-7 alkyl and C _3-8 cycloalkyl may optionally be substituted independently selected from up to six Substituents: halogen, cyano, _{nitro, SH, S (O) 0-2} R 5, NR 6 R 7, S (O) 2 NR 8 R 9, C (O) NR 10 R 11, C (O)OR ¹² , NR ¹³ S(O) ₂ R ¹⁴ , NR ¹⁵ C(O)R ¹⁶ , NR ¹⁷ C(O)OR ¹⁸ , NR ¹⁹ C(O)NR ²⁰ R ²¹ , OR ²² ), A benzene ring substituted with up to three substituents independently selected from the group consisting of halogen, cyano, nitro, SH, S(O) _0-2 R ⁵ , S(O) ₂ NR ⁸ R ⁹ , C ( O) NR ¹⁰ R ¹¹ , C(O)OR ¹² , OR ²² , C _1-6 alkyl or C _3-6 cycloalkyl, wherein the C _1-6 alkyl group and the C _3-6 cycloalkyl group may be as appropriate Substituted by up to three substituents independently selected from the group consisting of halogen, hydroxy, C _1-6 alkoxy, cyano, nitro, NH ₂ , NH(C _1-6 alkyl) and N (C _{1- 6} alkyl) ₂ ; R ⁵ , R ¹⁴ and R ¹⁸ represent C _1-6 alkyl or C _3-6 cycloalkyl optionally substituted by up to three substituents independently selected from halogen or hydroxy or C _1-6 alkoxy; R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² and R ²² each independently represent hydrogen or C _1-6 alkyl or C _3-6 cycloalkyl, wherein the C _{1 The 6} alkyl and C _3-6 cycloalkyl groups may be optionally substituted with up to three substituents independently selected from the group consisting of: , Hydroxy, C _1-6 alkoxy; or R ⁸ and R ⁹ or R ¹⁰ and R ¹¹ in any one group of connected together with the nitrogen atom may be taken together to form both the 4-6 aliphatic heterocycle, wherein the heteroaryl The ring may optionally be substituted with up to three substituents independently selected from the group consisting of halogen, hydroxy, _C1-6 alkoxy and _C1-6 alkyl; X represents O, S or S(O) ₂ .

m=0, 1, 2 or 3;

n=1, 2 or 3;

W represents CR ²⁷ R ²⁸ CR ²⁹ R ³⁰ or CR ³¹ R ³² CR ³³ R ³⁴ CR ³⁵ R ³⁶ .

V and Z independently represent the key, CR ³⁷ R ³⁸ or CR ³⁹ R ⁴⁰ CR ⁴¹ R ⁴² .

When X represents O, S or S(O) ₂ , the m, V and Z systems are such that all heteroatoms in the ring are separated by at least two carbon atoms (for example, when V is a bond, m is not 0, Z) Not for the key).

Y represents CO, CONR ⁴³ , and SO ₂ .

R ²⁷ , R ²⁸ , R ²⁹ , R ³⁰ , R ³¹ , R ³² , R ³³ , R ³⁴ , R ³⁵ , R ³⁶ , R ³⁷ , R ³⁸ R ³⁹ , R ⁴⁰ , R ⁴¹ and R ⁴² each independently represent hydrogen. , fluorine, C _1-6 alkyl or C _3-6 cycloalkyl.

When it does not represent hydrogen or fluorine, R ²⁷ and R ²⁸ , R ²⁹ and R ³⁰ , R ³¹ and R ³² , R ³³ and R ³⁴ , R ³⁵ and R ³⁶ , R ³⁷ and R ³⁸ , R ³⁹ and R ⁴⁰ Or R ⁴¹ and R ⁴² together with a carbon atom linking the two may form a 3 to 6 membered aliphatic ring; and R ⁴³ each independently represents hydrogen, C _1-6 alkyl or C _3-6 cycloalkyl. :L represents a linker comprising a straight or branched chain hydrocarbyl chain having up to 8 carbon atoms, such as 7, 6, 5 or 4 carbon atoms. Examples of such chaining include those of 4-7, 4-6, 4-5, 5-7, 6-7 and 5-6 carbon atoms.

L represents a C _4-8 alkyl chain optionally substituted with up to four (such as up to three, two or one) fluorine or methyl; suitably up to two carbon atoms of the chain may be independently selected from the following Substituting for the group: O, S, S(O) ₂ , NR ⁴⁶ C(O), C(O)NR ⁴⁷ , NR ⁴⁸ S(O) ₂ , S(O) ₂ NR ⁴⁹ , the limiting condition is In either case, any such group in the chain is separated by at least 2 carbon atoms; suitably up to four carbon atoms of the chain, such as up to three or up to two carbon atoms may form a portion of three monocyclic or bicyclic aromatic or heteroaromatic rings independently selected from heteroatoms of N, O or S, the ring comprising up to 10 ring atoms, and wherein the ring is optionally independently selected from up to three Substituted by the following substituents: halogen, S(O) _0-2 R ⁵⁶ , S(O) ₂ NR ⁵⁹ R ⁶⁰ , C(O)NR ⁶¹ R ⁶² , OR ⁷³ , C _1-6 alkyl and C _{3 a -6} cycloalkyl group, and wherein the C _1-6 alkyl group and the C _3-6 cycloalkyl group are optionally substituted with up to three substituents independently selected from the group consisting of halogen, hydroxy and C _1-6 alkoxy; Suitably the chain may comprise one such ring; suitably R ⁵⁶ represents C _1-4 alkane Or a C _3-6 cycloalkyl group, wherein the C _1-4 alkyl group and the C _3-6 cycloalkyl group are optionally substituted with up to three substituents independently selected from the group consisting of halogen, hydroxy, C _1-4 Alkoxy; and suitably R ⁴⁶ , R ⁴⁷ , R ⁴⁸ , R ⁴⁹ , R ⁵⁹ , R ⁶⁰ , R ⁶¹ , R ⁶² and R ⁷³ each independently represent hydrogen, C _1-4 alkyl or C _3-6 a cycloalkyl group, wherein the C _1-4 alkyl group and the C _3-6 cycloalkyl group are optionally substituted with up to three substituents independently selected from the group consisting of halogen, hydroxy, C _1-4 alkoxy; or R Any of ⁵⁹ and R ⁶⁰ or R ⁶¹ and R ⁶² together with the nitrogen atom linking the two may form a 4 to 6 membered aliphatic heterocyclic ring, wherein the heterocyclic ring may optionally be up to two independently selected from the group consisting of Substituted by a substituent: halogen, hydroxy, and C _1-4 alkyl or C _3-6 cycloalkyl, wherein the C _1-4 alkyl group and the C _3-6 cycloalkyl group are optionally independently selected from the group consisting of Substituted by a group of substituents: halogen, hydroxy and C _1-4 alkoxy; L ¹ , L ² , L ³ and L ⁴ independently represent hydrogen or methyl; R ¹ represents phenyl or has up to three independent a 5- to 6-membered heteroaryl ring from a hetero atom of N, O or S, in each case the ring Optionally substituted with up to three independently selected from the substituents: halogen, cyano, ^{nitro, C (O) OR 12,} OR 22, C 1-7 alkyl or C _3-8 cycloalkyl, wherein The C ₁ - ₇ alkyl group and the C _3-8 cycloalkyl group may be optionally substituted with up to three substituents independently selected from halogen, cyano, hydroxy or C _1-3 alkoxy; or R ¹ represents a fused aromatic or fused heteroaryl ring having up to 10 atoms and up to three heteroatoms independently selected from N, O or S, wherein in each case the ring may optionally be selected from up to three independently selected Substituted by a substituent: halogen, cyano, nitro, C(O)OR ¹² , OR ²² , C _1-6 alkyl or C _3-6 cycloalkyl, wherein the C _1-6 alkyl and C _{The 3-6} cycloalkyl group may be optionally substituted with up to three substituents independently selected from halogen, cyano, hydroxy or C _1-3 alkoxy; R ¹² and R ²² each independently represent hydrogen, C _{1- a 6} alkyl or C _3-6 cycloalkyl group, wherein the C _1-6 alkyl group or the C _3-6 cycloalkyl group may be optionally substituted with a substituent selected from the group consisting of halogen, hydroxy or C _1-3 alkane More suitable ring substituents include halogens such as fluorine and for example chlorine.

More suitable for the chemical type shown below - L-

Wherein ring D represents a phenyl, thiophene, furan or thiazole ring; R represents up to three ring substituents each independently selected from the group consisting of H, F, Cl, C _1-4 alkyl and CF ₃ ; t=0 or 1 ; G = O, CR ⁴³ R ⁴⁴ or S; when G = O or S, then v = 1 or 2; when G = C, then v = 0, 1 or 2, and wherein the chemical species - L- Attached to adjacent atoms in either orientation.

More suitable for the chemical species - L- is selected from - (phenyl-1,4-extended) OCH ₂ -, wherein the phenyl group is optionally substituted by 3, 2 or 1 methyl; - (phenyl-1, 4-extended) OCH ₂ CH ₂ -; -CH ₂ (benzene-1,4-extended) OCH ₂ -, wherein the phenyl group is optionally substituted by 3, 2 or 1 Cl or F (independent choice) ;-CH ₂ (benzene-1,4-exyl)-;CH ₂ (benzene-1,4-exyl)CH ₂ -;-(CH ₂ ) ₇ -; -CH ₂ (benzene-1,3- Exfoliation), wherein the benzene ring is optionally substituted with up to three C _1-3 alkyl groups, F, Cl and CF ₃ (independently selected); -CH ₂ (thiophene-2,5-extension)CH ₂ ----CH ₂ (thiophene-2,5-extension)-;-CH ₂ (thiophene-3,5-extension)-;-CH ₂ (thiophene-2,4-exetyl)-;-CH ₂ O (Benzene-1,3-Exo)-; and -CH ₂ S (Benzene-1,3-Exetyl)-.

Suitable linkers include such other :-( extending _{yl-1,5) - C (CH 3) 2-} ( phenyl-1,3-extending yl) - (benzene-1,3-extending yl) OCH ₂ CH ₂ --(Benzo-1,3-Exo)-C(CH ₃ ) ₂ -(CH ₂ ) ₄ --C(CH ₃ ) ₂ -(CH ₂ ) ₆ --CH ₂ (Benzene- 1,3-etyl) OCH ₂ -; -CH ₂ S(CH ₂ ) ₅ -; each of the foregoing linkers alone represents an independent aspect of the invention.

More suitable for the chemical species - L-system is selected from: -CH ₂ (thiophene-2,5-extension)-; -CH ₂ (benzene-1,3-exetyl)-, wherein the phenyl group can be singled by F Substituted; -CH ₂ (thiophene-2,4-exetyl)-; and -CH ₂ (thiophene-3,5-extension)-; more suitably L ¹ , L ² , L ³ and L ⁴ =H More suitably, the R ¹ is selected from any of the independent chemical species provided by the following examples.

X represents O or S.

m=1 or 2;

n=1 or 2;

W represents CR ²⁷ R ²⁸ CR ²⁹ R ³⁰ or CR ³¹ R ³² CR ³³ R ³⁴ CR ³⁵ R ³⁶ .

V and Z independently represent the key or CR ³⁷ R ³⁸ .

The V and Z systems are such that all heteroatoms in the ring are separated by at least two carbon atoms (for example, when V is a bond, the Z system is CR ³⁷ R ³⁸ ).

Y represents CO, CONR ⁴³ , SO ₂ ; such as CO or SO ₂ , such as CO.

R ²⁷ , R ²⁸ , R ²⁹ , R ³⁰ , R ³¹ , R ³² , R ³³ , R ³⁴ , R ³⁵ , R ³⁶ , R ³⁷ and R ³⁸ each independently represent hydrogen, fluorine or C _1-3 alkyl or C ₃ -cycloalkyl.

When it does not represent hydrogen or fluorine, R ²⁷ and R ²⁸ , R ²⁹ and R ³⁰ , R ³¹ and R ³² , R ³³ and R ³⁴ , R ³⁵ and R ³⁶ or R ³⁷ and R ³⁸ together with the carbon connecting the two The atoms together form a 3 to 6 membered aliphatic ring.

R ⁴³ each independently represents hydrogen or C _1-3 alkyl or C ₃ cycloalkyl is more preferably a bicyclic ring comprising -N-(^)mCVXWN(YR ¹ )-Z-(C)-(^)n- Covered by the spiral ring system:

(i) m and n=2, v=bond, Z=CH ₂ , X=O and W=CH ₂ CH ₂

(ii) m and n=2, v=bond, Z=CH ₂ , X=O and W=CF ₂ CH ₂

(iii) m and n=1, v=bond, Z=CH ₂ , X=O and W=CH ₂ CH ₂

(iv) m and n=2, v=bond, Z=CH ₂ CH ₂ , X=O and W=CH ₂ CH ₂

More preferably, the spiro ring system is selected from the foregoing (i) and (ii).

More suitably, the R ¹ group is selected from the group consisting of thiophene or thiazole or benzofuran, each optionally substituted with one or two substituents. One of the substituents optionally present is suitably selected from the group consisting of H, Cl, F and C _1-3 alkyl. Another substituent which is optionally present is selected from the group consisting of methyl, ethyl, propyl, butyl, CF ₃ , CH ₂ CF ₃ , CH(CH ₃ ) ₂ , CH(CH ₂ CH ₃ ) ₂ , CH(CH ₃ ) CH ₂ , CH ₃ , CH ₂ CH(CH ₃ ) ₂ , C(CH ₃ ) ₃ , cyclopropyl, cyclobutyl and cyclopentyl; each of the exemplified compounds of the invention represents a special and independent invention Aspect.

Suitable compounds of the invention include Examples 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12, 14, 15, 19, 22, 23, 25, 26, 27, 28, 29, 36, 37, 38, 40, 42, 44, 45, 47, 48, 52, 56, 57, 58, 66, 67, 70, 71, 75, 76, 82, 83, 84, 86, 87, 88, Compounds of 91, 93, 99, 105, 109, 111, 115, 265, 266, 278 and 280.

Suitable compounds of the invention include the compounds of Examples 22, 23, 36, 40, 42, 44, 47, 48, 57, 66, 82, 83, 84, 86, 87, 99, 265 and 266.

It will be appreciated that particular compounds of the invention may exist in a solvated (e.g., hydrated) and uncomplexed form. It is to be understood that the invention encompasses all such fused forms. A particular compound of formula (I) may exist in stereoisomeric form. It will be understood that the present invention encompasses all geometric and optical isomers of the compounds of formula (I) and mixtures thereof, including racemates. Tautomers and mixtures thereof also form aspects of the invention.

Suitably, the R-stereochemistry is possessed by a pair of palmar centers on a hydroxyl-substituted carbon atom adjacent to the beta-adrenergic receptor binding group.

It should also be understood that the present invention encompasses the replacement of any quaternary carbon with a ruthenium atom, in particular a quaternary carbon present in a spiro ring system, such as "Silicon switches of Marketed Drugs: Mini-reviews in Med. Chem.", 2006 , 6, 1169-1177.

In the context of the present specification, the term 'heteroaromatic' means a group comprising an optionally substituted aromatic monocyclic or polycyclic organic moiety having 5 to 14 ring atoms, preferably 5 to 10 ring atoms or A portion of a group in which up to four of the ring atoms are non-carbon elements such as nitrogen, oxygen or sulfur. Examples of such groups include benzimidazolyl, benzoxazolyl, benzothiazolyl, benzofuranyl, benzothienyl, furyl, imidazolyl, fluorenyl, pyridazinyl, sulphur Azyl, isoquinolyl, isothiazolyl, oxazolyl, oxadiazolyl, pyrazinyl, Zinyl, pyrazolyl, pyridyl, pyrimidinyl, pyrrolyl, quinolinolinyl, quinolinyl, tetrapyridyl, 1,3,4-thiadiazolyl, thiazolyl, thienyl and triazolyl. The heteroaryl group can be substituted with one or more substituents. A heteroaryl group can be attached to the remainder of the compounds of the invention by any available carbon or nitrogen atom.

The term 'aliphatic heterocyclic ring' means a non-aromatic ring comprising at least one hetero atom selected from the group consisting of nitrogen, oxygen and sulfur. Examples of the 4-8 membered aliphatic heterocyclic ring of the present invention include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, homopiperazinyl, homopiperidinyl and azetidinyl.

Unless otherwise stated, in the context of the present specification, alkyl groups and moieties may be straight or branched and include, for example, methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso - butyl or tert-butyl. The cycloalkyl group is a monocyclic ring such as a cyclopentyl group or a cyclohexyl group. The halogen is, for example, a fluorine, a chloride or a bromide.

In the context of the present specification, when the stated group may optionally be substituted with up to three substituents, the group may be unsubstituted or substituted; when substituted, the group is usually substituted by one, two or three Substituted by the base. Typically, the hydroxyl moiety is not attached to a carbon atom adjacent to a nitrogen atom, another oxygen atom or a sulfur atom.

The invention further provides a process for the preparation of a compound of formula (I), or a pharmaceutically acceptable salt thereof, as hereinbefore defined, which comprises:

(a) when L ¹ represents hydrogen, the compound of formula (II) or a suitable salt thereof

Wherein LG ¹ represents a leaving group (eg, chloride, bromide, iodide, mesylate or p-toluenesulfonate) and L, L ² , L ³ , L ⁴ , R ¹ , m, n, V, W, X, Y and Z are as defined in formula (I), and a compound of formula (III) or a suitable salt thereof (for example, hydrobromide, acetate or hydrochloride),

Wherein Ar in the system, such as formula (I) defined above, and P ¹ is hydrogen or a protecting group (e.g., the third - butyldimethyl silicon group) in the presence of a base (e.g. potassium carbonate, triethylamine or diisopropylethylamine Ethylamine) reaction, followed by removal of the protecting group (for example using hydrofluoric acid-pyridine complex); or

(b) when L ¹ represents hydrogen, the compound of formula (IV) or a suitable salt thereof

Wherein L, L ² , L ³ , L ⁴ , R ¹ , m, n, V, W, X, Y and Z are as defined in formula (I), and a compound of formula (III) or a suitable salt thereof is suitably a reducing agent such as sodium cyanoborohydride, sodium triethoxysulfonate or hydrogen in the presence of palladium or platinum oxide catalyst on a suitable carbon; or

(c) a compound of formula (V) or a suitable salt thereof,

Wherein L, L ¹ , L ² , L ³ , L ⁴ , R ¹ , m, n, V, W, X, Y and Z are as defined in formula (I), and P ³ represents hydrogen or an activating group (eg 3-nitrophenylsulfonyl) and a compound of formula (VI) or a suitable salt thereof,

Wherein Ar is as defined in formula (I), LG ² represents a leaving group (eg, chloride, bromide, iodide, mesylate or p-toluenesulfonate) and P ¹ is as defined in formula (III); the presence of a base (e.g. when P ³ is hydrogen system potassium carbonate, triethylamine, diisopropyl ethyl amine, and, when P ³ is 3-nitrophenyl based sulfo acyl as sodium hydride or diisobutyl Lithium propylamine) reaction, followed by removal of a protecting group (for example using hydrofluoric acid-pyridine complex, thiophenol, thioacetic acid); or with a compound of formula (VII) or a suitable salt thereof,

Wherein Ar is as defined in formula (I); in the presence of a base (for example, when P ³ is hydrogen, potassium carbonate, triethylamine, diisopropylethylamine, and when P ³ is 3-nitro) a phenylsulfonyl group for sodium hydride or lithium diisopropylamide), followed by removal of a protecting group (eg, trifluoroacetic acid, thiophenol, sulfuric acid); or with a compound of formula (VIII) or a suitable salt thereof ,

Wherein Ar is as defined in formula (I), and LG ² represents a leaving group (eg, chloride, bromide, iodide, mesylate or p -toluenesulfonate) in the presence of a base (eg, when the P ³ system is hydrogen) a reaction of potassium carbonate, triethylamine, diisopropylethylamine, and when the P ³ is a 3-nitrophenylsulfonyl group, sodium hydride or lithium diisopropylamide, followed by the ketone Reduction (for example using sodium borohydride or borane / palmitic catalyst complex) followed by removal of protecting groups (eg trifluoroacetic acid, thiophenol, sulfuric acid); or

(d) when each of L ³ and L ⁴ represents hydrogen, the compound of formula (IX) or a suitable salt thereof,

Wherein Ar, L, L ¹ and L ² are as defined in formula (I), P ¹ is as defined in formula (III), and P ³ represents a protecting group (for example, a third -butyl urethane or 3- Nitrophenylsulfonyl); and a compound of formula (X) or a suitable salt thereof,

Wherein R ¹ , m, n, V, W, X, Y and Z are as defined in formula (I) in the presence of a suitable reducing agent (eg sodium cyanoborohydride, sodium triethoxy borohydride or Reaction of hydrogen in the presence of palladium or platinum oxide catalyst on a suitable carbon, followed by removal of the protecting group (for example, treatment with hydrochloric acid or trifluoroacetic acid thiophenol, thioacetic acid); or

(e) when one or both of L ³ and L ⁴ represents hydrogen, the compound of formula (XI) or a suitable salt thereof,

Wherein Ar, L, L ¹ and L ² are as defined in formula (I), P ¹ is as defined in formula (III), and P ³ represents a protecting group (eg, tert-butylcarbonyl or 3-nitrophenyl) Sulfhydryl), LG ³ represents a leaving group (eg, chloride, bromide, iodide, mesylate or p -toluenesulfonate), and a compound of formula (X) or a suitable salt thereof, in the presence of a base (eg Reaction with potassium carbonate, triethylamine, diisopropylethylamine, followed by removal of a protecting group (eg, trifluoroacetic acid, thiophenol, sulfuric acid); or

(f) when each of L ¹ and L ² represents hydrogen, the compound of formula (XII) or a suitable salt thereof, wherein Ar, L, L ³ , L ⁴ , m, n, V, W, X, Y and Z formula-based definition (the I), P ¹ and system as in formula (III) defined above, and P ⁴ based nitrogen protecting group of the appropriate (e.g., tertiary - butyl carbonate); and of a suitable reducing agent (e.g., boron Alkane tetrahydrofuran complex), followed by removal of a protecting group (for example using a hydrofluoric acid-pyridine complex) and reaction with a compound of formula (XIII) or a suitable salt thereof,

Wherein R ¹ and Y are as defined in formula (I), and LG ⁴ represents a hydroxy group or a cleavage group (eg, halide, chloride) or a suitable salt thereof, followed by removal of a protecting group (eg, using hydrofluoric acid-pyridine pyridine) Compound, hydrochloric acid or trifluoroacetic acid).

When LG ⁴ represents a hydroxyl group, the reaction is suitably an activator (for example, carbonyldiimidazole, 1-propanephosphonic acid cyclic anhydride (T3P) or hexafluorophosphate O-(7-azabenzotriazol-1-yl) In the presence of -N,N,N',N'-tetramethyluronium (HATU), in an organic solvent (eg N,N -dimethylformamide or dichloromethane), optionally in the presence of a base In the presence of (e.g., triethylamine), for example, at a temperature ranging from 0 to 60 ° C, and when LG ⁴ represents a halide (e.g., chloride), the reaction is suitably a base (e.g., triethylamine, diisopropyl) In the presence of a benzylamine or a pyridine, in an organic solvent such as dichloromethane or tetrahydrofuran, for example at a temperature in the range of from 0 to 25 ° C; and optionally in (a), (b), (c) And (d), (e) or (f) are followed by one or more of the following steps: converting the resulting compound to another compound of the invention to form a pharmaceutically acceptable salt of the compound.

In the methods (a), (c) and (e), the reaction may suitably be carried out in an organic solvent such as N,N -dimethylformamide, ethanol, n-butanol or dimethylarylene. It is carried out, for example, at a temperature in the range of 50 to 140 °C.

In methods (b) and (d), the reaction may suitably be carried out in an organic solvent containing up to 10% by weight of water and acetic acid (such as methanol, ethanol, dichloromethane, acetic acid, N -methylpyrrolidone or N, It is carried out in N -dimethylformamide.

In the process (f), the reaction can be suitably carried out in an organic solvent such as tetrahydrofuran at a temperature in the range of, for example, 0 to 80 °C.

The compound of the formula (II) can be obtained by using a compound of the formula (XIV) or a suitable salt thereof.

Wherein L, L ³ , L ⁴ , R ¹ , m, n, V, W, X, Y and Z are as defined in formula (II), and a compound of formula (XV)

Wherein L ² is as defined in formula (II) and Mt represents a metal such as lithium or magnesium or aluminum or boron (eg methyl lithium, methyl magnesium bromide, lithium aluminum hydride, sodium borohydride); in organic solvents (eg tetrahydrofuran or diethyl ether), the reaction was carried out at a temperature range of 0 to 60 deg.] C of example, and then a hydroxyl group formed of transforming into suitable of leaving group (e.g. chloride, bromide, iodide, mesylate or p - toluene sulfonamide Prepared by acid).

The compound of the formula (IV) can be reacted with a compound of the formula (XIV) and a compound of the formula (XV) in an organic solvent (for example, tetrahydrofuran or diethyl ether) at a temperature in the range of, for example, -10 to 60 ° C, followed by an organic solvent (such as In methylene chloride, N , N -dimethylformamide or dimethyl hydrazine, at a temperature ranging, for example, from -78 to 60 ° C, with a suitable oxidizing agent (eg Swern reagent, Dess-Martin reagent or It is prepared by oxidizing the hydroxy group formed by pyridinium chlorochromate.

a compound of the formula (V), wherein L ¹ represents hydrogen and L, L ² , L ³ , L ⁴ , P ³ , R ¹ , m, n, V, W, X, Y and Z are as defined in the formula (V), Can be prepared by the following method

a) reacting a compound of the formula (II) with sodium azide in an organic solvent such as tetrahydrofuran, N , N -dimethylformamide or dimethylhydrazine in a temperature range of, for example, 25 to 85 ° C, The formed azide compound is then reduced in an organic solvent (eg, tetrahydrofuran and water) using a suitable reducing agent (eg, triphenylphosphine), and finally the amine formed is protected (eg, 3-nitrobenzene in the presence of a base such as pyridine) Sulfonyl chloride treatment); or

b) reacting a compound of formula (IV) with an amine such as benzylamine, alpha-methylbenzylamine, 4-methoxybenzylamine or 2,4-methoxybenzylamine, followed by Use an appropriate reducing agent (for example, 10% by weight of water and an organic solvent of acetic acid such as methanol, ethanol, dichloromethane, acetic acid, N -methylpyrrolidone or N , N -dimethylformamide) Reduction of the imine formed by sodium cyanoborohydride or sodium triethoxysulfonate, followed by an organic solvent (eg ethanol, methanol, tetrahydrofuran, dichloromethane, acetonitrile, water or a mixture thereof) at 25 to 80 Use appropriate reagents at temperatures up to °C (eg hydrogen and appropriate catalysts (palladium or palladium hydroxide on carbon), 2,3-dichloro-5,6-dicyanobenzoindole (DDQ) or ammonium cerium nitrate (CAN)) removal of the benzyl protecting group formed, followed by protection of the amine formed (for example, treatment with 3-nitrophenylsulfonyl chloride in the presence of a base such as pyridine); a compound of formula (V) wherein L, L ¹ , L ² , L ³ , L ⁴ , P ³ , R ¹ , m, n, V, W, X, Y and Z are as defined in formula (V), by making a compound of formula (XVI) or suitable Salt,

Wherein LG ⁵ is a leaving group (for example, hydroxyl or chlorine), and L, L ¹ , L ² , L ³ , L ⁴ , R ¹ , m, n, V, W, X, Y and Z are as in formula (V). defined in the following reagent was prepared with, such as when the system when LG ⁵ is hydroxyl, in the presence of an amine (e.g. triethylamine) in an organic solvent (e.g. III - butanol, tetrahydrofuran, dichloromethane, water or mixtures thereof In the range of 25 to 100 ° C, with diphenylphosphonic acid azide, or when LG ⁵ is chlorine, in an organic solvent (such as diethyl ether, tert -butanol, tetrahydrofuran, water or its In the mixture), it is reacted with sodium azide at a temperature ranging from 25 to 100 ° C (Angewandte Chemie, 2005, 54, 5188), and finally, the amine formed is subsequently protected (for example, 3-nitrate in the presence of a base such as pyridine) Phenyl sulfonium chloride treatment).

Compounds of formula (III), (VI), (VII), (VIII), (XIII) and (XV) are known in the literature or can be prepared using known techniques.

The compound of formula (IX) can be prepared by the following method

a) a compound of formula (XVII) or a suitable salt thereof,

Wherein P ⁵ is hydrogen or a protecting group (for example, tert-butyldimethylmethylalkyl, tetrahydropyran) and P ³ , L, L ¹ and L ² are as defined in formula (IX), and are present in the base. (e.g. when P ³ is hydrogen system potassium carbonate, triethylamine or diisopropyl ethylamine, sodium hydride and based or diisopropylethylamine when P ³ is 3-nitrophenyl based sulfo acyl Lithium alkoxide) in an organic solvent such as N,N-dimethylformamide, N-methylpyrrolidone, tetrahydrofuran, ethanol, n-butanol or dimethylarylene, for example 50 to 140 The compound of formula (VI), (VII) or (VIII) or a suitable salt thereof is reacted at a temperature in the range of °C. When the reaction is carried out with a compound of formula (VIII), a second step involving reduction of the ketone (e.g., using sodium borohydride or borane/p-capacity catalyst complex) is required. Properly selective removal of the protecting group (eg hydrofluoric acid-pyridine complex, tetrabutylammonium fluoride, dilute hydrochloric acid or amberlyst-15 resin in methanol) with the appropriate oxidizing agent (pyridinium chlorochromate, Dess -Martin reagent or Swern reagent) oxidizing the formed alcohol to the corresponding aldehyde to produce a compound of formula (IX); or

b) a compound of formula (VIII) or a suitable salt thereof,

Wherein P ⁶ and P ⁷ represent an acyclic or cyclic carbonyl protecting group (eg, dimethoxy or diethoxy acetal, 1,3-dioxopentane or 1,3-dioxane) and L, L ¹ , L ² , and P ³ are as defined in formula (IX), in the presence of a base (for example, when the P ³ system is hydrogen, potassium carbonate, triethylamine or diisopropylethylamine, and when P ³ is a sodium hydride or lithium diisopropylamide in the case of 3-nitrophenylsulfonyl), in an organic solvent (such as N,N-dimethylformamide, N-methylpyrrolidone, The tetrahydrofuran, ethanol, n-butanol or dimethylhydrazine is reacted with a compound of the formula (VI), (VII) or (VIII) or a suitable salt thereof at a temperature of, for example, 50 to 140 °C. When reacted with a compound of formula (VIII), this is followed by reduction of the ketone (e.g., using sodium borohydride or borane/pivotic catalyst complex). Removing a protecting group (eg, dilute hydrochloric acid or amberlyst-15 resin in methanol) to produce a compound of formula (IX); or

c) when L ¹ represents hydrogen, the compound of formula (XIX) or a suitable salt thereof,

Wherein P ⁵ is hydrogen or a protecting group (eg, tert-butyldimethylmethylalkyl, tetrahydropyran) and L and L ² are as defined in formula (IX), in the presence of a suitable reducing agent (eg, Sodium cyanoborohydride, sodium triethoxy borohydride or hydrogen in the presence of palladium or platinum oxide catalyst on a suitable carbon) in an organic solvent (such as methanol, containing up to 10% by weight of water and acetic acid) Ethyl alcohol, methylene chloride, acetic acid, N -methylpyrrolidone or N , N -dimethylformamide, reacted with a compound of formula (III) or a suitable salt thereof, followed by appropriate selective removal protection Base (eg hydrofluoric acid-pyridine complex, tetrabutylammonium fluoride, dilute hydrochloric acid or amberlyst-15 resin in methanol) with a suitable oxidizing agent (pyridinium chlorochromate, Dess-Martin reagent or Swern reagent) Oxidizing the formed alcohol to the corresponding aldehyde; or

d) when L ¹ represents hydrogen, the compound of formula (XX) or a suitable salt thereof,

Wherein P ⁶ and P ⁷ represent an acyclic or cyclic carbonyl protecting group (eg, dimethoxy or diethoxy acetal, 1,3-dioxopentane or 1,3-dioxane) and L and L ² is as defined in formula (IX) in the presence of a suitable reducing agent (for example, sodium cyanoborohydride, sodium triethoxy borohydride or hydrogen in the presence of palladium or platinum oxide catalyst on a suitable carbon) ) in an organic solvent (such as methanol, ethanol, dichloromethane, acetic acid, N -methylpyrrolidone or N , N -dimethylformamide) containing up to 10% by weight of water and acetic acid, The compound of formula (III) or a suitable salt thereof is reacted, followed by removal of a protecting group (e.g., dilute hydrochloric acid or amberlyst-15 resin in methanol).

The compound of the formula (XI) can be produced by converting a compound of the formula (IX) or a precursor into a compound of the formula (IX) as described above, and selecting an appropriate reaction sequence, for example, reducing an aldehyde to an alcohol (for example, sodium borohydride), suitably Selective removal of a protecting group (eg, hydrofluoric acid-pyridine complex, tetrabutylammonium fluoride, dilute hydrochloric acid or amberlyst-15 resin in methanol) and conversion of the alcohol to a suitable cleavage group (eg, chloride, Bromide, iodide, mesylate or p -toluenesulfonate; or

The compound of the formula (XII) can be obtained by using a compound of the formula (XXI) or a suitable salt thereof.

Wherein L, L ³ , L ⁴ , P ⁴ , m, n, V, W, X and Z are as defined in formula (XII) and LG ⁶ represents a hydroxyl group or a leaving group (for example, chlorine), and a compound of formula (III) or a salt of an appropriate; when LG ⁶ represents hydroxyl, the reaction system is suitably an activator (e.g., carbonyl diimidazole, 1-propanephosphonic acid cyclic anhydride (T3P) hexafluorophosphate or O- (7- aza-benzene In the presence of triazol-1-yl)-N,N,N',N'-tetramethyluronium (HATU) in an organic solvent (eg N , N -dimethylformamide or dichloromethane) In the case of a base (for example, triethylamine), for example, at a temperature in the range of 0 to 60 ° C, and when LG ⁶ represents chlorine, the reaction is suitably a base (for example, triethylamine or diiso) In the presence of propylethylamine), it is carried out in an organic solvent such as dichloromethane or tetrahydrofuran at a temperature in the range of, for example, 0 to 25 °C.

Compounds of formula (V), (X), (XIV), (XVI) and (XXI) may be via a compound of formula (XIII) wherein R ¹ , Y are as defined in formula (I), and LG ⁴ represents hydroxy or detachment The base (e.g., halide, chloride) or a suitable salt thereof is obtained by a general coupling reaction. When LG ⁴ represents a hydroxyl group, the reaction is suitably an activator (e.g., carbonyldiimidazole, 1-propanephosphonic acid cyclic anhydride). (T3P) or hexafluorophosphate O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium (HATU) in the presence of an organic solvent ( For example, N , N -dimethylformamide or dichloromethane), optionally in the presence of a base such as triethylamine, is carried out, for example, at a temperature ranging from 0 to 60 ° C, or when LG ⁴ represents a halogen a root (eg, a chloride), suitably in the presence of a base such as triethylamine, diisopropylethylamine or pyridine in an organic solvent such as dichloromethane or tetrahydrofuran, for example 0 to Using a compound of the formula (XXII) or a suitable salt thereof at a temperature in the range of 25 ° C,

Wherein m, n, V, W, X, Y and Z are as defined in formula (I) and - in the case of the compound of formula (V), P ⁹ represents

Wherein L, L ¹ , L ² , L ³ , L ⁴ and P ³ are as defined in formula (V); and in the case of the compound of formula (X), P ⁹ represents a suitable nitrogen protecting group, such as a third -butyl group. Oxycarbonyl, or - in the case of a compound of formula (XIV), P ⁹ represents

Wherein L, L ³ and L ⁴ are as defined in formula (XIV), wherein P ¹¹ and P ¹² represent an acyclic or cyclic carbonyl protecting group (eg dimethoxy or diethoxy acetal, 1,3- Dioxopentane or 1,3-dioxane) followed by appropriate deprotection (eg dilute hydrochloric acid or amberlyst-15 resin in methanol); - in the case of the compound of formula (XVI), P ⁹ represents

Wherein L, L ¹ , L ² , L ³ and L ⁴ are as defined in formula (XVI), wherein P ¹⁴ represents an acid protecting group (eg methyl, ethyl or tert-butyl), followed by appropriate deprotection (e.g., lithium hydroxide or sodium hydroxide, trifluoroacetic acid, hydrochloric acid); - in the case of the compound of formula (XXI), P ⁹ represents

Wherein L, L ³ and L ⁴ are as defined in formula (XXI), wherein P ¹⁴ represents an acid protecting group (eg, a third-butyl group), followed by appropriate deprotection (eg, trifluoroacetic acid, hydrochloric acid); XXII) a compound wherein V represents a bond, X represents O, W represents CR ²⁷ R ²⁸ CR ²⁹ R ³⁰ , Z represents CR ³⁷ R ³⁸ , and R ²⁷ , R ²⁸ , R ²⁹ , R ³⁰ , R ³⁷ , R ³⁸ represent Hydrogen and P ⁹ represents a suitable nitrogen protecting group such as a third -butoxycarbonyl group, which can be prepared from a compound of (XXIII)

Wherein P ⁹ , m and n are as defined in the compound of formula (XXII) by treatment with a suitable reducing agent such as a borane-THF complex at 30-70 ° C in a suitable solvent such as tetrahydrofuran. The formed boron complex is decomposed in methanol at 60 to 90 ° C with an appropriate amine such as N1,N 2 -dimethyleneamine-1,2-diamine

Compounds of formula (XXIII) can be prepared from compounds of formula (XXIV)

Wherein LG ⁷ is a suitable leaving group, such as a halogen or tosylate and P ⁹ , m and n are as defined in the compound of formula (XXIII), which is in a suitable solvent such as tetrahydrofuran at 50 to 90 ° C. A suitable base such as potassium t-butoxide is used.

The compound of the formula (XXIV) can be produced by reacting a compound of the formula (XXV) with a compound of the formula (XXVI).

Wherein LG ⁸ represents a hydroxyl group or a halogen group, such as a chloride group, and P ⁹ , m, n and LG ⁷ are as defined in the compound of the formula (XXIV); when LG ⁸ represents a hydroxyl group, the reaction is suitably an activator (for example, a carbonyl group). Diimidazole, 1-propanephosphonic acid cyclic anhydride (T3P) or hexafluorophosphate O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium ( In the presence of HATU)) in an organic solvent (for example N , N -dimethylformamide or dichloromethane), optionally in the presence of a base such as triethylamine, for example in the range from 0 to 60 °C Performing at a temperature; when LG ⁸ represents a halide (for example, a chloride), the reaction is suitably carried out in the presence of a base such as triethylamine, diisopropylethylamine or pyridine in an organic solvent such as dichloromethane. Or tetrahydrofuran), for example, at a temperature ranging from 0 to 25 ° C;

A compound of formula (XXV) can be obtained by formulating a compound of formula (XXVII)

Wherein P ⁹ , m and n are as defined in the compound of formula (XXV), and are reacted with ammonia in a suitable solvent such as methanol at a temperature in the range of from 20 to 60 ° C;

A compound of formula (XXVII) can be obtained by formulating a compound of formula (XXVIII)

Wherein P ⁹ , m and n are as defined in the compound of formula (XXVII), in the presence of a suitable base such as sodium hydride or potassium butoxide, in a suitable solvent such as dimethyl hydrazine at 0 to 20 ° C Prepared by reaction with trimethylammonium iodide at a temperature within the range; the above method also represents a simple oxidation and reduction step, which is carried out under standard conditions well documented in the literature (eg Dess-Martin, Swern) , pyridinium chlorochromate, sulfur trioxide pyridinium complex oxidation). It can be suitably carried out in an organic solvent such as dichloromethane at a temperature ranging from -78 to 50 °C (Annual Reports on the Progress of Chemistry, Section B: Organic Chemistry, 2004, 100 , 51-70).

Compounds of formula (VI), (VII), (VIII), (XIII), (XV), (XVII), (XIX), (XX), (XXVI) and (XXVIII) are commercially available, known or may be Those skilled in the art can readily prepare using one of the foregoing methods or using known techniques.

Other intermediate compounds are novel and represent independent aspects of the invention.

It is known to those skilled in the art that in the methods of the invention, specific functional groups in the reagent may need to be protected by a protecting group. Thus, the preparation of the compound of formula (I) provides for the addition or removal of one or more protecting groups at an appropriate stage.

Functional group protection and deprotection are discussed in JWF McOmie's 'Protective Groups in Organic Chemistry', Plenum Press (1973) and 'Protective Groups in Organic Synthesis', 3 ^rd edition, TW Greene and PGM Wuts, Wiley-Interscience ( 1999).

The compound of formula (I) can be converted to another compound of formula (I) using standard methods.

The compounds of formula I have activity as pharmaceuticals, especially as dual adrenergic beta agonists and anticholinergic agents, including muscarinic receptor (M1, M2 and M3) antagonists, especially M3 antagonists. Diseases and conditions which can be treated using a compound of formula (I) and a pharmaceutically acceptable salt thereof include:

1. Respiratory tract: obstructive diseases of the respiratory tract, including: asthma, including bronchi, allergic, endogenous, exogenous, exercise-induced, drug-induced (including aspirin and NSAID) and dust-induced asthma, including intermittent and Persistence and all other causes of severity and respiratory hyperactivity; chronic obstructive pulmonary disease (COPD); bronchitis, including infectious and eosinophilic bronchitis; emphysema; bronchiectasis; cyst fibrosis; Sarcoma disease; peasant lung and related diseases; allergic pneumonia; pulmonary fibrosis, including cryptogenic fibrotic alveolitis, idiopathic interstitial pneumonia, anti-tumor therapy with fibrosis and chronic infection, including tuberculosis and sputum Diseases and other fungal infections; complications of lung transplantation; vasculitis of the pulmonary vessels and embolic disorders, and pulmonary hypertension; anti-tissue activity includes treatment of chronic cough associated with respiratory tract inflammation and secretion, and iatrogenic cough Acute and chronic rhinitis including drug-induced rhinitis and vasomotor rhinitis; long-term and seasonal allergic rhinitis, including neuropathic rhinitis (pollen Hot); nasal meat; acute viral infections, including general colds and infections caused by respiratory fused cell viruses, influenza, coronavirus (including SARS) or adenovirus; or eosinophilic esophagitis;

2. Bones and joints: joint diseases associated with or including osteoarthritis/non-inflammatory osteoarthrosis, including primary and secondary, such as congenital hip dysplasia; cervical and lumbar disc disease and lower back And neck pain; osteoporosis; rheumatoid arthritis and Still's disease; seronegative spondyloarthropathy, including ankylosing spondylitis, joint psoriasis, reactive arthritis, and undifferentiated spine Arthropathy; septic arthritis and other infection-related joint and bone disorders, such as tuberculosis, including Potts'disease and Poncet's syndrome; acute and chronic crystallization caused by synovitis , including uric acid gout, calcium pyrophosphate deposition disease and apatite-related tendons, bursae and synovial inflammation; Behcet's disease; primary and secondary Sjogren's syndrome Systemic sclerosis and localized scleroderma; systemic lupus erythematosus, mixed-type tissue disease and undifferentiated tissue disease; inflammatory myopathy, including dermatomyositis and polymyositis; Sexual polyarthritis; juvenile arthritis, including idiopathic inflammatory joint disease and associated syndromes and rheumatic fever and systemic complications wherever joints are distributed; vasculitis, including giant cell arteritis, high arteritis ( Takayasu's arteritis), Churg-Strauss syndrome, nodular polyarteritis, microscopic polyarteritis, and vasculitis associated with viral infections, allergic reactions, psychrophilic globulins and paraproteins; lower back pain; familial Mediterranean fever, Muckle-Wells syndrome and familial Irish fever, Kikuchi disease; drug-induced joint pain, tendinitis and myopathy;

3. Pain in musculoskeletal conditions due to injury [eg sports injuries] or disease and association of tissue reconstruction: arthritis (eg rheumatoid arthritis, osteoarthritis, gout or crystalline joint disease), other joint diseases (such as Conical disc degeneration or temporomandibular joint degeneration), bone remodeling diseases (such as osteoporosis, Paget's disease or osteonecrosis), polychondritis, scleroderma, mixed connective tissue disorders, spinal joints Lesions or periodontal disease (such as periodontitis);

4. Skin: psoriasis, atopic dermatitis, contact dermatitis or other eczema dermatitis and delayed allergic reactions; plant- and photodermatitis; seborrheic dermatitis, herpes-like dermatitis, flat Moss, sclerosing atrophic moss, gangrenous pyoderma, skin sarcoidosis, discoid lupus erythematosus, pemphigus, pemphigoid, bullous epidermolysis, urticaria, angioedema, vasculitis, Toxic erythema, cutaneous eosinophilia, alopecia areata alopecia, male regular alopecia, Sweet's syndrome, Weber-Christian syndrome, polymorphic erythema; cellulitis, both infectious and non-infectious; panniculitis Skin lymphoma, non-melanoma skin cancer and other adverse lesions; drug-induced conditions, including fixed drug eruptions;

5. Eyes: eyelid inflammation; conjunctivitis, including long-term and spring-type allergic conjunctivitis; iritis; anterior and posterior uveitis; choroiditis; autoimmune; affecting the degenerative or inflammatory condition of the omentum; ocular inflammation, including Sensible ophthalmia; sarcoma-like disease; infection, including viruses, fungi and bacteria;

6. Gastrointestinal tract: Candida type glossitis, gingivitis, periodontitis; esophagitis, including reflux; eosinophilic gastroenteritis, mastocytosis, Crohn's disease, colitis including ulcerative colitis, Proctitis, anal pruritus; abdominal disease, colonic irritability, and food-related allergies (such as migraine, rhinitis, or eczema) that may have little effect on the gut;

7. Abdomen: hepatitis, including autoimmune, alcohol and viruses; fibrosis and cirrhosis of the liver; cholecystitis; pancreatitis, both acute and chronic;

8. Genitourinary organs: nephritis, including interstitial and glomerulonephritis; renal syndrome; cystitis, including acute and chronic (interstitial) cystitis and Hunner's ulcer; acute and chronic urethra Inflammation, prostatitis, parastatitis, oophoritis and salpingitis; vulvovaginitis; Peyronie's disease; erectile dysfunction (both men and women);

9. Implant rejection: acute and chronic, such as after kidney, heart, liver, lung, bone marrow, skin or corneal transplantation or after transfusion; or chronic graft versus host disease;

10. CNS: Alzheimer's disease and other dementia disorders, including CJD and nvCJD; amyloidosis; multiple sclerosis and other demyelinating syndromes; cerebral atherosclerosis and vasculitis; Inflammation; myasthenia gravis; acute and chronic pain (severe, intermittent or persistent, central or peripheral sources) including visceral pain, headache, migraine, trigeminal neuralgia, atypical dermatological pain, joint and bone pain, from cancer and cancer Invasive pain, neuropathic pain syndrome, including diabetic, postherpetic and HIV-related neuropathy; neurosarcoma; malignant, infectious or autoimmune process center and peripheral nervous system complications;

11. Other autoimmune and allergic conditions, including Hashimoto's thyroiditis, Graves' disease, Addison's disease, diabetes, idiopathic thrombocytopenic purpura, eosinophils Membrane inflammation, IgE hypersensitivity syndrome, antiphospholipid syndrome;

12. Other conditions with inflammatory or immune components; including acquired immunodeficiency syndrome (AIDS), rickets, Sezary syndrome, and paraneoplastic syndrome;

13. Cardiovascular: atheroma hardening, affecting coronary and peripheral circulation; pericarditis; myocarditis, inflammatory and autoimmune cardiomyopathy, including myocardial sarcoidosis; ischemia reperfusion injury; endocarditis, Heart valvular inflammation and aortitis, including infectious (eg syphilis); vasculitis; conditions of proximal and peripheral veins, including phlebitis and embolism, including complications of deep vein thrombosis and varicose veins;

14. Tumor: treatment of general cancer, including prostate, breast, lung, ovary, pancreas, intestine and colon, stomach, skin and brain tumors and malignant diseases affecting bone marrow (including leukemia) and lymphoproliferative system, how are Jay's (Hodgkin's and non-Hodgkin's lymphoma; including prevention and treatment of metastatic disease and tumor recurrence, and paraneoplastic syndrome; and

15. Gastrointestinal tract: abdominal disease, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease, ulcerative colitis, microscopic colitis, undetermined colitis, irritable bowel syndrome, colonic urgency, Non-inflammatory diarrhea, food-related allergies (such as migraine, rhinitis, or eczema) that may have little effect on the bowel.

Accordingly, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, for use in therapy as defined hereinbefore.

In another aspect, the invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, as hereinbefore defined, for the manufacture of a medicament for use in therapy.

In the context of the present specification, the term "treatment" also includes "prevention" unless specifically stated to the contrary. The terms "therapeutic" and "prophylactic" should be as described.

It is expected that prevention will be particularly relevant to treatment in the early stages of the disease or condition being studied or as an increased risk of illness. Those at risk of developing a particular disease or condition typically include a family history of the disease or condition or have been genetically tested or screened to determine a particularly prone to develop the disease or condition.

The invention still further provides a method of treating or reducing the risk of an inflammatory disease or condition, including a reversible obstructive respiratory disease or condition, comprising administering a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, as hereinbefore defined Apply to patients in need.

In particular, the compounds of the invention are useful in the treatment of adult dyspnea syndrome (ARDS), emphysema, bronchitis, bronchiectasis, chronic obstructive pulmonary disease (COPD), asthma, and rhinitis.

For the purposes of the aforementioned therapeutic use, the dosage administered will of course vary depending on the compound employed, the mode of administration, the treatment desired, and the condition to be treated. For example, the daily dose of the compound of the invention may range from 0.05 micrograms per kilogram body weight (μg/kg) to 100 micrograms per kilogram body weight (μg/kg) upon inhalation. Alternatively, if the compound is administered orally, the daily dose of the compound of the invention may range from 0.01 micrograms/kg body weight ([mu]g/kg) to 100 mg/kg body weight (mg/kg).

The compound of the formula (I) and pharmaceutically acceptable salts thereof may be used alone, but are usually administered in the form of a pharmaceutical composition wherein the compound (salt) of the formula (I) (active ingredient) is pharmaceutically acceptable adjuvant, diluent or Carrier binding. Conventional methods for selecting and preparing suitable pharmaceutical formulations are described, for example, in "Pharmaceuticals-The Science of Dosage Form Designs", M. E. Aulton, CHurchill Livingstone, 1988.

The pharmaceutical composition preferably comprises from 0.05 to 99% by weight, more preferably from 0.05 to 80% by weight, still more preferably from 0.10 to 70% by weight, still more preferably from 0.10 to 50% by weight, of the active ingredient, depending on the mode of administration. All weight percentages are based on the overall composition.

The invention also provides a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, as hereinbefore defined, in association with a pharmaceutically acceptable adjuvant, diluent or carrier.

The invention further provides a process for the preparation of a pharmaceutical composition of the invention comprising admixing a compound of formula (I) as hereinbefore defined, or a pharmaceutically acceptable salt thereof, with a pharmaceutically acceptable adjuvant, diluent or carrier.

Pharmaceutical compositions may, for example, be in the form of creams, solutions, suspensions, hydrofluoroalkane (HFA) aerosols, and dry powder formulations, for example The formulation of the inhalation device is administered topically (for example, to the skin or to the lungs and/or the respiratory tract); or by systemic administration by oral administration, for example, in the form of a tablet, capsule, syrup, powder or granule; or in the form of a solution or suspension. Parenteral administration; or subcutaneous administration; or rectal administration as a suppository; or transdermal administration.

Dry powder formulations of the compounds of the invention and pressurized aerosols can be administered by oral or nasal inhalation. When inhaled, it is desirable that the compound be in the form of a fine powder. The finely powdered compound preferably has a mass median diameter of less than 10 μm, and may be by means of a dispersing agent such as a C ₈ -C ₂₀ fatty acid or a salt thereof (for example, oleic acid), a bile salt, a phospholipid, an alkyl sugar, a perfluorinated or A polyethoxylated surfactant or other pharmaceutically acceptable dispersing agent is suspended in the propellant mixture.

The compounds of the invention may also be administered by dry powder inhalers. The inhaler can be a single or multiple dose inhaler and can be a breath activated dry powder inhaler.

One possibility is to mix the finely divided compounds of the invention with carrier compounds such as monosaccharides, disaccharides or polysaccharides, sugar alcohols or other polyols. Suitable carriers are sugars such as lactose, glucose, plant honey, raffinose, lactitol, maltitol, saccharose, sucrose, mannitol, and starch. Or the finely powdered compound can be coated with other substances. The powder mixture can also be dispensed into hard gelatin capsules, each containing the desired amount of active compound.

Another possibility is to process the finely powdered powder into a ball which breaks during inhalation. The spheroidized powder can be filled into a drug container of a multi-dose inhaler, for example Wherein the dosage unit measures the required dose, which is subsequently inhaled by the patient. Using such a system, the active ingredient (along with or without carrier material) is delivered to the patient.

When administered orally, the compound of the present invention may be combined with an adjuvant or carrier such as lactose, sucrose, sorbitol, mannitol; starch, such as potato starch, corn starch or starch pectin; cellulose derivative; binder, For example, gelatin or polyvinylpyrrolidone; and/or a lubricant such as magnesium stearate, calcium stearate, polyethylene glycol, wax, paraffin, and the like, followed by compression. If an ingot is desired, the core prepared as described above may be coated with a concentrated sugar solution, which may contain, for example, gum arabic, gelatin, talc, and titanium dioxide. Alternatively, the tablet may be coated with a suitable polymer dissolved in a volatile organic solvent.

In the preparation of soft gelatin capsules, the compounds of the invention may be admixed, for example, with vegetable oils or polyethylene glycols. The hard gelatin capsules may contain the particles of the compound using the excipients mentioned above for the tablet. Liquid or semi-solid formulations of the compounds of the invention may also be filled into hard gelatin capsules.

The liquid preparation for oral administration may be in the form of a syrup or suspension, for example, a solution containing the compound of the present invention, the balance being a mixture of sugar and ethanol, water, glycerin and propylene glycol. Such liquid preparations may optionally contain coloring agents, flavoring agents, saccharin and/or carboxymethylcellulose as a thickening agent or other excipients known in the art.

In particular, the compounds of the invention and their salts are useful in the treatment of inflammatory diseases such as, but not limited to, rheumatoid arthritis, osteoarthritis, asthma, allergic rhinitis, chronic obstructive pulmonary disease (COPD), psoriasis and inflammatory bowel disease The compounds of the invention may be combined with a non-steroidal anti-inflammatory agent (hereinafter referred to as NSAID), including a non-selective cyclooxygenase COX-1/COX-2 inhibitor (whether locally or systemically) (such as Piroxicam, diclofenac, propionic acid such as naproxen, flurbiprofen, fenoprofen, ketoprofen and ibuprofen (ibuprofen), fenamate derivatives such as mefenamic acid, indomethacin, sulindac, azapropazone, pyrazolone ) such as phenylbutazone, salicylate such as aspirin; selective COX-2 inhibitors (such as meloxicam, celecoxib, rofecoxib) ), valdecoxib, lumarocoxib, parecoxib, and ito Ethoicoxib; a cyclooxygenase (CINOD) that inhibits nitric oxide donors; glucocorticoids (whether administered by local, oral, intramuscular, intravenous or intra-articular routes); aminomethyl folate; Leflunomide; hydroxychloroquine; d-penicillamine; auranofin or other parenteral or oral gold preparations; analgesics; diacerein; intra-articular agents such as hyaluronic acid a derivative; and a nutritional supplement such as glucosamine.

The compounds of the invention may also be administered concurrently with other compounds useful in the treatment of the aforementioned conditions.

The invention therefore further relates to combination therapies wherein the compound of the invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or formulation comprising a compound of the invention is administered simultaneously or sequentially or in combination with other therapeutic or therapeutic agents The dosage form is administered to treat one or more of the foregoing conditions.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof, together with a cytokine or an agonist or antagonist having cytokine function (including agents acting on a cytokine message delivery pathway, such as modulators of the SOCS system) Including α-, β- and γ-interferon; insulin-like growth factor type I (IGF-1); interleukolin (IL) including IL1 to 17, and an interleukin antagonist or inhibitor, such as ana Anakina; tumor necrosis factor alpha (TNF-alpha) inhibitors, such as anti-TNF monoclonal antibodies (eg, infliximab; adalimumab and CDP-870) and TNF receptor antagonists, including immunoglobulin molecules (such as etanercept) and low molecular weight agents, such as a combination of pentoxyfylline.

Further, the present invention relates to a compound of the present invention or a pharmaceutically acceptable salt thereof and a labeled B-lymphocyte (such as CD20 (rituximab), MRA-aIL16R) or T-lymphocytes (CTLA4-Ig, HuMax) Il-15) A combination of monoclonal antibodies.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof and a modulator having a chemotactic hormone receptor function such as CCR1, CCR2, CCR2A, CCR2B, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCR10 and CCR11 (CC family); a combination of CXCR1, CXCR2, CXCR3, CXCR4 and CXCR5 (CXC family) and CX ₃ CR1 f or an antagonist of the CX ₃ -C family.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof and an inhibitor of matrix metalloproteinase (MMP), namely matrix lysin, collagenase and gelatinase, and aggrecanase; in particular collagenase-1 (MMP-1) ), collagenase-2 (MMP-8), collagenase-3 (MMP-13), matrix lysin-1 (MMP-3), matrix lysin-2 (MMP-10), and matrix lysin-3 ( MMP-11) and MMP-9 and MMP-12 include combinations of agents such as doxycycline.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof and a leukotriene biosynthesis inhibitor, a 5-lipoxygenase (5-LO) inhibitor or a 5-lipoxygenase activating protein (FLAP) Antagonists such as; zileuton; ABT-761; fenleuton; tepoxalin; Abbott-79175; Abbott-85761; N-(5-substituted)-thiophene-2 - alkyl sulfonamide; 2,6-di-tertiary-butyl phenolphthalein; methoxytetrahydropyran, such as zeneca ZD-2138; compound SB-210661; pyridinyl substituted 2-cyanonaphthalene A compound such as L-739, 010; a 2-cyanoquinoline compound such as L-746, 530; or a quinone or quinoline compound such as a combination of MK-591, MK-886 and BAY x 1005.

The invention further relates to a combination of a compound of the invention or a pharmaceutically acceptable salt thereof and an acceptor antagonist of leukotrienes (LT) B4, LTC4, LTD4 and LTE4, selected from the group consisting of phenothiazine-3- 1, such as L-651,392; mercapto compounds, such as CGS-25019c; benzoxazole amines, such as ontazolast; benzamidine, such as BIIL 284/260; and compounds such as zafirlukast ), albukast, montelukast, pranlukast, verlukast (MK-679), RG-12525, Ro-245913, Iras Iralukast (CGP45715A) and BAY×7195.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof and a phosphodiesterase (PDE) inhibitor such as methylxanthanine, including theophylline and the amine theophylline; a selective PDE isomeric isomerase Inhibitors, including combinations of PDE4 inhibitors, inhibitors of the isoform PDE4D, or inhibitors of PDE5.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof and a histamine type 1 receptor antagonist such as cetirizine, loratadine, desloratadine, Fexofenadine, acrivastine, terfenadine, astemizole, azelastine, levocabastine, chlorpheniramine (chlorpheniramine), a combination of promethazine, cyclizine or mizolastine; administered orally, topically or parenterally.

The invention further relates to a combination of a compound of the invention or a pharmaceutically acceptable salt thereof and a proton pump inhibitor (such as omeprazole) or a gastric protective histamine type 2 receptor antagonist.

The invention further relates to a combination of a compound of the invention or a pharmaceutically acceptable salt thereof and an antagonist of a histamine type 4 receptor.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof and an α-1/α-2 adrenergic receptor agonist vasoconstrictory sympathomimetic agent such as propylhexedrine, phenylephrine , phenylpropanolamine, ephedrine, pseudoephedrine, nepyrimazine hydrochloride, hydroxyisoxazoline hydrochloride, tetrahydrozoline hydrochloride, xylometazoline hydrochloride A combination of a salt, tramazoline hydrochloride or ethyl norepinephrine hydrochloride.

The invention further relates to a compound of the invention, or a pharmaceutically acceptable salt thereof, and a chromone, such as a combination of sodium cromoglycate or nedocromil sodium.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof and a glucocorticoid such as flunisolide, triamcinolone acetonide, beclomethasone dipropionate, budesonide A combination of (budesonide), fluticasone propionate, ciclesonide or mometasone furoate.

The invention further relates to a combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and a medicament for modulating a nuclear hormone receptor, such as PPAR.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof, together with an immunoglobulin (Ig) or Ig preparation or an antagonist or antibody which modulates Ig function, such as anti-IgE (e.g., omalizumab) combination.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof and to another systemically or topically applied anti-inflammatory agent such as thalidomide or a derivative thereof, a retinoid, a dithranol or A combination of calcipotriol.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof and an aminosalicylate and a sulfapyridine such as sulfasalazine, mesalazine, alsalarazide and olsalazine ( Olsalazine); and combinations of immunomodulatory agents such as thiopurine and a combination of corticosteroids such as budesonide.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof together with an antibacterial agent such as a penicillin derivative, a tetracycline, a macrolide, a beta-indoleamine, a fluoroquinolone, an imidazodazole (metronidazole), inhaled aminoglycoside; antiviral agents include acyclovir, famciclovir, valaciclovir, ganciclovir, cidofovir, Amantadine, rimantadine, ribavirin, zanamavir and oseltamavir; protease inhibitors such as indinavir, naphthalene Nelfinavir, ritonavir and saquinavir; nucleoside reverse transcriptase inhibitors such as didanosine, lamivudine, sitaf Stavudine, zalcitabine or zidovudine; or a combination of non-nucleoside reverse transcriptase inhibitors such as nevirapine or efavirenz.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof and a cardiovascular agent such as a calcium channel blocker, a beta-adrenergic receptor blocker, an angiotensin-converting enzyme (ACE) inhibitor, vasoconstriction a receptor-2 receptor antagonist; a lipid lowering agent such as statin or a fibric acid derivative; a blood cell type modulator such as pentoxyfylline; a thrombolytic or anticoagulant such as a platelet aggregation inhibitor The combination.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof and a CNS agent such as an antidepressant (such as sertraline), an anti-Parkinson drug (such as deprenyl, L-dopa) , ropinirole (ropinipole), pramipexole, MAOB inhibitors such as selegine and rasagiline, comP inhibitors such as tasmar, A-2 inhibitors , dopamine recovery inhibitors, NMDA antagonists, nicotine agonists, dopamine agonists or inhibitors of neuronal nitric oxide synthase) or anti-Alzheimer's drugs such as donepezil (depezil), rivastigmine ( Combination of rivastigmine), tacrine, COX-2 inhibitor, propentofylline or metrifonate.

The invention further relates to a compound of the invention, or a pharmaceutically acceptable salt thereof, and an agent for the treatment of acute or chronic pain, such as a central or peripheral acting analgesic (such as opium or a derivative thereof), carbamazepine, A combination of phenytoin, sodium valproate, amitryptiline or other anti-depressant, paracetamol or a non-steroidal anti-inflammatory agent.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof in combination with a parenteral or topical (including inhalation) local anesthetic such as lignocaine or a derivative thereof.

The compound of the present invention or a pharmaceutically acceptable salt thereof can also be used in combination with an anti-osteoporosis agent including a hormonal agent such as raloxifene or a hydrogenphosphonate such as alendronate.

The invention further relates to a compound of the invention or a pharmaceutically acceptable salt thereof, together with: (i) a neutral protease inhibitor; (ii) a platelet activating factor (PAF) antagonist; (iii) a neutral interleukin converting enzyme (ICE) Inhibitors; (iv) IMPDH inhibitors; (v) adhesion molecule inhibitors including VLA-4 antagonists; (vi) cathepsins; (vii) kinase inhibitors such as tyrosine kinase inhibitors (such as Btk, Itk, Jak3 or MAP, such as Gefitinib or Imatinib mesylate), serine/threonine kinase (such as MAP kinase inhibitors such as p38, JNK, protein kinase A, B or C or IKK) or a kinase involved in cell cycle regulation (such as a cyclin-dependent kinase); (viii) a glucose-6 phosphate dehydrogenase inhibitor; (ix) kinin-B.sub1- or B.sub2.- Receptor antagonist; (x) anti-gout agent, such as colchicine; (xi) inhibitor such as allopurinol; (xii) promotes uric acid excretion, such as probenecid, sulfonate Sulfonepyrazone or benzbromarone; (xiii growth hormone secretagogue; (xiv) transforming growth factor (TGFβ); (xv) platelet-derived Growth factor (PDGF); (xvi) fibroblast growth factor such as basic fibroblast growth factor (bFGF); (xvii) granule/macrophage colony-stimulating factor (GM-CSF); (xviii) (xix) agonistic peptide NK.sub1. or NK.sub3. receptor antagonists such as NKP-608C, SB-233412 (talnetant) or D-4418; (xx) elastase inhibitors such as UT-77 or ZD-0892; (xxi) TNF-α converting enzyme inhibitor (TACE); (xxii) induces one nitric oxide synthase (iNOS) inhibitor; (xxiii) chemokine receptor-homology expressed on TH2 cells Molecules, (such as CRTH2 antagonists); (xxiv) inhibitors of P38; (xxv) agents that regulate receptor-like receptor (TLR) function, (xxvi) agents that regulate receptor activity such as P2X7; (xxvii) transcription factors A combination of an inhibitor of activation such as NFkB, API or STATS; or (xxviii) a glucocorticoid receptor (GR-receptor) agonist.

In another aspect, the invention provides a compound of formula (I) and one or more combinations selected from the group consisting of the following: (eg, for the treatment of COPD, asthma, or allergic rhinitis): a non-steroidal glucocorticoid receptor (GR-receiver) agonist; ● PDE4 inhibitor, including inhibitor of isoform PDE4D; ● modulator of chemotactic hormone receptor function (such as CCR1 receptor antagonist); ● steroid (such as budesonide) Budesonide; and ● inhibitor of p38 kinase function.

The compound of the present invention or a pharmaceutically acceptable salt thereof can also be used in combination with an existing therapeutic agent for treating cancer, for example, including the following agents: (i) an anti-cell proliferation/anti-tumor drug or a combination thereof, which is used in oncology, Such as alkylating agents (eg cisplatin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan, chloram bucil, busulphan or nitrosourea) Antimetabolite (eg antifolate, such as fluoropyrimidine, such as 5-fluorouracil or tegafur, raltitrexed, aminomethylfolate, cytarabine, hydroxyurea, gemcitabine) (gemcitabine) or paclitaxel); anti-tumor antibiotics (eg, anthracyclines such as doxorubicin, bleomycin, doxorubicin, daunomycin, epirubicin) ), idarubicin, mitomycin-C, dactinomycin or mithramycin; anti-mitotic agents (eg vinblastine, such as vincristine, vinblastine) , vindesine or vinorelbine or yew (taxoid), such as taxol or taxotere; or topoisomerase inhibitors (eg, epitopophyllotoxin, such as etoposide, teniposide) (teniposide), amsacrine, topotecan or camptothecin; (ii) cytostatic agents such as antiestrogens (eg tamoxifen, torre) Toremifene, raloxifene, droloxifene or iodoxyfene, estrogen receptor modulator (eg fulvestrant), antibiotic Androgens (such as bicalutamide, flutamide, nilutamide or cyproterone acetate), LHRH antagonists or LHRH agonists (eg goserelin) (goserelin), leuprorelin or buserelin, progesterone (eg medroxyprogesterone acetate), aromatase inhibitors (eg anastrozole, letrozole) (letrozole), vorazole or exemestane) or an inhibitor of 5α-reductase, such as finasteride; (iii) An agent that targets cancer cell invasion (eg, a metalloproteinase inhibitor such as marimastat or an inhibitor of urokinase-type plasmin activating factor function); (iv) a growth factor function inhibitor, such as a growth factor antibody ( For example, the anti-erbb2 antibody trastuzumab or the anti-erbb1 antibody cetuximab [C225], a farnesyl transferase inhibitor, a tyrosine kinase inhibitor or a serine/ A sulphate kinase inhibitor, an epidermal growth factor family inhibitor (eg, an EGFR family tyrosine kinase inhibitor such as N- (3-chloro-4-fluorophenyl)-7-methoxy-6-(3- Morpholinylpropoxy)quinazolin-4-amine (gefitinib, AZD1839), N- (3-ethynylphenyl)-6,7-bis(2-methoxyethoxy Or quinazolin-4-amine (erlotinib, OSI-774) or 6-propenyl decylamino- N- (3-chloro-4-fluorophenyl)-7-(3- Morpholinylpropoxy)quinazolin-4-amine (CI 1033), an inhibitor of the platelet-derived growth factor family or an inhibitor of the hepatocyte growth factor family; (v) an anti-angiogenic agent, such as a blood vessel The role of endothelial growth factor (eg anti-vascular endothelial cells) A long-term factor antibody, bevacizumab, a compound disclosed in WO 97/22596, WO 97/30035, WO 97/32856, or WO 98/13354, or a compound that acts by another mechanism (eg, tris-carboxyaminoquinoline ( Linomide), an inhibitor of integrin αvβ3 function or angiostatin; (vi) a vascular disrupting agent, such as combestatin A4 or WO 99/02166, WO 00/40529, WO 00/ a compound disclosed in 41669, WO 01/92224, WO 02/04434 or WO 02/08213; (Vii) an agent for antisense therapy, such as one of the targets listed above, such as ISIS 2503, anti- Ras antisense; (Viii) agents for use in gene therapy methods, such as methods for replacing aberrant genes, such as aberrant p53 or abnormal BRCA1 or BRCA2, GDEPT (gene-directed enzyme prodrug-therapeutic) methods, such as the use of cytosine Deaminase, thymidine kinase or bacterial nitroreductase, and methods for increasing tolerance of a patient to chemotherapy or radiation therapy, such as multi-drug resistance gene therapy; or (ix) for immunotherapeutic methods An agent, such as a body that increases the immunity of a patient's tumor cells -and- in vivo methods, such as transfection with cytokines such as interleukin 2, interleukin 4 or granulocyte-macrophage colony stimulating factor, to reduce the unresponsive state of T-cells, using transfection Immunocytic cells such as cytokine-transfected dendritic cells, methods using cytokine-transfected tumor cell lines, and anti-idiotypic antibodies are used.

The aforementioned compound of formula (I) can be converted into a pharmaceutically acceptable salt thereof, such as an acid addition salt such as a hydrochloride (e.g., a dihydrochloride), a hydrobromide (e.g., a dihydrobromide), trifluoro Acetate (eg di-trifluoroacetate), sulfate, phosphate, acetate, fumarate, maleate, tartrate, lactate, citrate, pyruvate, grass An acid salt, a methanesulfonate or a p-toluenesulfonate.

The invention is illustrated by the following examples without restricting the invention, in which the following general methods are used:

General method

¹ H NMR spectra were recorded on a Varian Inova 400 MHz or Varian Mercury -VX 300 MHz instrument. Using a central peak of chloroform- d (δ _H 7.27 ppm), dimethyl sulfonium- d ₆ (δ _H 2.50 ppm), acetonitrile- d ₃ (δ _H 1.95 ppm) or methanol- d ₄ (δ _H 3.31 ppm) As an internal reference. Column chromatography was performed using tannin: Fisher Scientific silica 60A, particle size 35-70 microns, Or 0.040-0.63mm, pre-filled biotage KP-Sil column. Starting materials are commercially available unless otherwise stated. All solvents and commercially available reagents are laboratory grade and can be used in the form obtained.

LC/MS analysis uses the following methods:

Instrument Agilent 1100; column Waters Symmetry C ₁₈ , 2.1 × 50 mm; mass spectrometry APCI or multimode (APCl + ESI; flow rate 1 mL / min; wavelength 220 nm; solvent A: water + 0.1% TFA; solvent B: acetonitrile + 0.1% TFA ; 5 to 95%/B gradient in 8 minutes.

Reverse-phase preparative HPLC system used in aqueous 0.2% TFA or 0.1% aqueous formic acid gradient of acetonitrile or methanol, using SunFire ^TM prep C8OBD ^TM 5μm30 × 100mm column (Waters Corporation), a flow rate of 35mL / min, or 0.1% Gradient of acetonitrile or methanol in aqueous ammonium acetate or 0.1% aqueous formic acid 50 x 19 mm column (Waters Corporation), flow rate 18.5 mL/min.

General Methods LC/MS Analysis for Examples 101 to 115 and 279 to 285 The following method was used: the final compound was analyzed using MS3 and the intermediate was used MS4 MS3: instrument Waters Micromass ZQ quadrupole mass spectrometer connected to a Hewlett Packard HP1100 LC system. Sample injection was performed by a Gilson 215 autosampler. The spectrometer has an electrical spray source that operates in both positive and negative ion modes. Additional detection is achieved using the Sedex 55 Evaporative Light Scattering Detector. Flow rate 1 ml/min; wavelength 254 nm; solvent A: water + 0.1% formic acid; solvent B: acetonitrile + 0.1% formic acid; gradient from 5 to 95% B for 20 min. MS4: connected to a detector with UV diode array and automatic The sampler's Hewlett Packard 1050 LC system instrument Finnigan AQ single quadrupole mass spectrometer. The spectrometer has an electrical spray source that operates in positive ion mode.

Additional detection is achieved using the Sedex 65 Evaporative Light Scattering Detector. Flow rate 1 ml/min; wavelength 254 nm; solvent A: water + 0.1% formic acid; solvent B: acetonitrile + 0.1% formic acid; gradient from 5 to 95% B over 5 min.

NMR spectroscopy is recorded on one of three instruments: the Varian Unity Inova 400 spectrometer, ¹ H operating at 400 MHz, and the device is useful for detecting ¹ H, ¹³ C, ³¹ P 5 mm inverse detection triple resonance probes, with 9.4 The magnetic field provided by the Tesla Oxford instrument superconducting magnet and the Sun Microsystems SunBlade 1000 workstation.

Bruker Avance DRX 400 spectrometer, ¹ H 400MHz in operation, the device is useful to detect ¹ H, ¹³ C, ¹⁵ N of 5mm inverse detection triple resonance probe TX1, having a magnetic field provided by a 9.4 Tesla Oxford instruments as a superconducting magnet, and The main computer is an HP workstation wx5000 operated by WIN-NMR software under Windows XP.

Bruker Avance DPX 300 spectrometer, 1H operating at 300MHz, the device is used to detect 1H and 13C standard 5mm dual-frequency probe, with magnetic field provided by 7.05Tesla Bruker superconducting magnet and as the main body under Windows 2000 with Bruker XWIN- HP workstation for NMR software operation.

Column chromatography was performed using silicone: Fluka Silicone 60, 35-70 μm particle size, pre-filled Teledyne Isco, Inc. RediSep Rf column or pre-filled Isolute Flash Si II SPE column. All solvents and commercially available reagents are laboratory grade and can be used in the form obtained.

Reverse phase preparative HPLC purification was performed using a gradient of acetonitrile in 0.1% aqueous TFA or 0.1% aqueous formic acid using Phenomenex Gemini A C18 column (250 x 21.2 mm, 5 microns) was run as a static phase at a flow rate of 18 mL/min.

The abbreviations or terms used in the examples have the following meanings: SCX: strong cation exchange - sulfonic acid adsorbent using cerium oxide as the main solid phase extraction HPLC: high performance liquid chromatography DCM: dichloromethane DMF: N, N -dimethylformamide NMP: 1-methylpyrrolidin-2-one THF: tetrahydrofuran TFA: trifluoroacetic acid DMSO: dimethyl hydrazine aq: aqueous solution h: hour min: minute g: gram mL: ml RT: room temperature MP-TsOH65: Macroporous polymer-bonded ion exchange resin HATU hexafluorophosphate O- (7-azabenzotriazol-1-yl) -N,N,N', provided by Biotage N' -tetramethyluronium hydrazine NBS N -bromosuccinimide T3P propanephosphonic anhydride TBAF tetrabutylammonium fluoride CDI 1,1'-carbonyldiimidazole MTBE methyl third -butyl ether MCPBA inter-chlorine benzoic acid

Varian bonded dissociation NH ₂ column: strong anion exchange. Solid Phase Extraction with Oxide as NH2 Adsorbent 1-Oxo-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester hydrochloride was purchased from WuXi Pharma Tech 7-[(1 R )-2-Amino-1-hydroxy-ethyl]-4-hydroxy-3H-benzothiazol-2-one acetate or HCl salt (WO 2007027134, Example 1, step d ) is 86 to 94% ee(R)-5-(2-amino-1-(t-butyldimethyl decyloxy)ethyl)-8-hydroxyquinoline-2 (1H) -ketone (WO 2004106333) is 92 to 96% ee

Named set software for the title compound/subtitle compound:

Struct=Cam bridgeSoft Corporation name 9.0.7

Example 1 ( R )-4-hydroxy-7-(1-hydroxy-2-(4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)ethoxy)benzylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 7-(2-Chloro-ethenyl)-4-hydroxy-3H-benzothiazol-2-one

Ethanol (1500 mL) was added to 7-acetamido-4-hydroxy-3H-benzothiazol-2-one (150 g) (WO 2004/016578) and dichloroiodic acid in a flask equipped with an overhead stirrer. In a mixture of benzyltrimethylammonium (374 g). The mixture was heated to 78 ° C for 1 h and allowed to cool to room temperature overnight. The mixture was poured into water (2 L), and the precipitate was collected by filtration, washed with water, filtered to dryness, and suspended in ethyl acetate. The mixture was heated to reflux and allowed to cool to room temperature with stirring. The solid was collected by filtration, washed with cold ethyl acetate (200 mL) and then taken to diethyl ether (1L). The solid was collected, washed with EtOAc (EtOAc)EtOAc. The yield was 164 g.

m/z 244(M+H) ⁺ (APCI)

b) 7-(2-azido-ethenyl)-4-hydroxy-3H-benzothiazol-2-one

7-(2-Chloro-ethenyl)-4-hydroxy-3H-benzothiazol-2-one (Example 1, step a) (331 g) was dissolved in N,N -dimethylformamide (1800 mL) And stirred in an ice bath for 10 minutes. Sodium azide (88.3 g) was added in portions over 15 minutes. The reaction was stirred for 72 hours, then the reaction mixture was divided into three equal portions and the reaction was quenched in ice and water (2.5 L). The solid was filtered off and washed with water (1 L) and resuspended in acetonitrile (1.5 L). The solvent was evaporated and another portion of acetonitrile (1 L) was added and the solvent was evaporated again to dry. Diethyl ether (1.5 L) was added and the mixture was stirred to give a homogeneous suspension. The solid was collected and dried under vacuum at 35 ° C for 24 hours to give sub-title compound. The yield was 285 g.

m/z 251(M+H) ⁺ (APCI)

c) 7-[(1 R )-2-azido-1-hydroxy-ethyl]-4-hydroxy-3H-benzothiazol-2-one

(1 R , 2 S )-(+)-cis-1-amino-2-nonanol (149 g) was added in portions at 25 to 25 ° C for 25 minutes to the borane-tetrahydrofuran complex ( 1M in tetrahydrofuran, 2997 mL). The mixture was stirred at 20 ° C for an additional 30 minutes, cooled to 0 ° C, and 7-(2-azido-ethenyl)-4-hydroxy-3H-benzothiazole-2- was added in portions at a temperature maintained at 0 to 5 °C. Ketone (Example 1, step b) (250 g). The reaction mixture was stirred at 0 &lt;0&gt;C for additional 1 h and then quenched with methanol (350 g, 442 <RTIgt; The exotherm allowed the temperature to reach 17 °C. The reaction was evaporated to a brown foam and redissolved in ethyl acetate (1.2L). Aqueous hydrochloric acid (87 mL cone. HCl in 1.2 L water) was added and the mixture was stirred vigorously for 30 min. The aqueous layer was separated and washed with fresh ethyl acetate (2×600 mL). The combined organic solution was washed with water (1.2 L). The aqueous layer was filtered through EtOAc (EtOAc) (EtOAc) Combined ethyl acetate solution was dried in Na ₂ SO _4, filtered and evaporated. The resulting solid was suspended in 5% EtOAc / EtOAc (EtOAc) The yield was 213 g.

m/z 253 (M+H) ⁺ (APCI)

d) 7-[(1R)-2-Amino-1-hydroxy-ethyl]-4-hydroxy-3H-benzothiazol-2-one, acetate

Palladium-type 87 L slurry (22 g) on 5% carbon was added to 7-[(1 R )-2-azido-1-hydroxy-ethyl]-4-hydroxy-3H- dissolved in ethanol (3 L). Benzothiazol-2-one (Example 1, step c) (225 g). The reaction was stirred under hydrogen (3 bar) for 48 h. An additional 10 g of catalyst was added and hydrogenation was continued for another 5 days. The reaction mixture was filtered and the solid (product + catalyst) was suspended in ethanol (2.5 L), then acetic acid (150 mL) was added and stirred overnight. The mixture was filtered again to remove the palladium catalyst on the carbon. The solution was evaporated to dryness and azeotroped with toluene (2×1L). The solid was stirred in tetrahydrofuran (1 L) for 4 h, filtered and dried <RTI ID=0.0> The yield was 57 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) .δ6.85 (d, 1H), 6.69 (d, 1H), 4.54 (dd, 1H), 2.78-2.67 (m, 2H) +5 exchangeable protons

e) 4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester

1-Propanephosphonic acid cyclic anhydride (1.57 M solution in THF, 4.18 mL) was added to 1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester salt A solution of the acid salt (WuXi PharmaTech) (1.92 g), 2-methylthiazole-4-carboxylic acid (0.94 g) and triethylamine (5.48 mL) in DMF (70 mL). The reaction mixture was poured into water (500 mL) The combined organic solution was washed with water (2×100 mL) and brine (100 mL). Purification by gelatin chromatography eluting with ethyl acetate gave the title compound as a clear oil. The yield was 2.30 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ7.95 (s, 1H), 3.80-3.45 (m, 8H), 3.18-2.96 (m, 2H), 2.67 (s, 3H), 1.77- 1.62 (m, 2H), 1.43-1.31 (m, 11H)

f) (2-Methylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecethane-4-yl)methanone hydrochloride

Trifluoroacetic acid (10 mL) was added to 4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tertidine The ester (Example 1, step e) (2.3 g) in DCM (50 mL) in EtOAc. The solvent was evaporated in vacuo. Toluene (50 mL) was added and the mixture was evaporated in vacuo. The residue was dissolved in methanol (20 mL) and applied to EtOAc EtOAc. The column was washed with methanol (250 mL) and dissolved (3 mL) with 3M aqueous methanol. The eluent was evaporated in vacuo and the residue was dissolved in MeCN (100 mL). Add HCl (1M in EtOAc (EtOAc)MeOHMeOH The yield was 1.90 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 9.16 (s, 2H), 7.92 (s, 1H), 4.25 (s, 4H), 3.66-3.58 (m, 2H), 3.12-2.90 (m) , 4H), 2.69 (s, 3H), 2.01-1.89 (m, 2H), 1.85-1.73 (m, 2H).

g) [4-(2,2-Diethoxy-ethoxy)-phenyl]-methanol

Cesium carbonate (39.4 g) was added to a mixture of 4-hydroxymethyl-phenol (10 g) and 2-bromo-1,1-diethoxyethane (12.73 mL) in DMF (200 mL). The mixture was stirred at 90 ° C for 16 h. The reaction was poured into water (500 mL) andEtOAcEtOAc The combined organic solution was washed with water (250 mL) and brine (250 mL). Purification by gel chromatography on a gradient of isohexane to diethyl ether afforded the title compound as a yellow oil. The yield was 9.5 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.29-7.25 (m, 2H), 6.93-6.88 (m, 2H), 4.83 (t, J = 5.2Hz, 1H), 4.61 (s, 2H), 4.00 ( d, J = 5.2 Hz, 2H), 3.81-3.72 (m, 2H), 3.68-3.58 (m, 2H), 1.25 (t, J = 7.0 Hz, 6H). An unobserved exchangeable proton.

h) 2-(4-(hydroxymethyl)phenoxy)acetaldehyde

2M HCl (4 mL) was added to a solution of (4-(2,2-diethoxyethoxy)phenyl)methanol (Example 1, step g) (0.9 g) in EtOAc (20 mL) The resulting mixture was stirred at room temperature for 16 h. The reaction was concentrated in vacuo and aq. EtOAc (EtOAc) The combined organic solution was dried with MgSO4, filtered and evaporated in vacuo. The yield was 0.50 g.

i) (9-(2-(4-(hydroxymethyl)phenoxy)ethyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)( 2-methylthiazol-4-yl)methanone

(2-Methylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone hydrochloride (Example 1, step f (0.38 g) added to a solution of 2-(4-(hydroxymethyl)phenoxy)acetaldehyde (Example 1, step h) (0.17 g) in NMP (10 mL) and acetic acid (0.06 mL) . The resulting mixture was stirred for 30 min and then cooled in an ice bath. Sodium triethoxy borohydride (0.32 g) was then added and the reaction allowed to warm to rt and stirred 16 h. The reaction was diluted with methanol (30 mL) and applied to an SCX column pre-wetted with methanol. The column was washed with methanol (250 mL) and dissolved (3 mL) with 3M aqueous methanol. The eluent was evaporated in vacuo and the residue was purified eluting eluting elut elut elut The yield was 0.32 g.

_{^{1 H NMR (300MHz, D 6}} -DMSO) δ7.86 (s, 1H), 7.23-7.16 (m, 2H), 6.85 (dt, J = 8.7,1.1Hz, 2H), 4.72-4.62 (m, 1H ), 4.44 - 4.38 (m, 2H), 4.10-3.99 (m, 2H), 3.66 (d, J = 6.7 Hz, 4H), 3.61-3.55 (m, 2H), 2.71-2.64 (m, 5H), 2.47-2.42 (m, 4H), 1.76-1.64 (m, 2H), 1.59-1.45 (m, 2H).

j) 4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethoxylate Benzaldehyde

Manganese dioxide (0.65g) was added to (9-(2-(4-(hydroxymethyl)phenoxy)ethyl)-1-oxa-4,9-diazaspiro[5.5] eleven Carboxane-4-yl)(2-methylthiazol-4-yl)methanone (0.32 g) (Example 1, step i) in DCM (20 mL) Heat down for 1 h. After cooling, the reaction mixture was passed through a pad of Celite. The pad was washed with DCM (2 x 30 mL) and the combined filtrate and washings were evaporated in vacuo to give a sub-title compound. The yield is 0.25 g.

m/z 430 (M+H) ⁺ (APCI) ¹ H NMR (300MHz, D _{6 -} DMSO) δ 9.86 (s, 1H), 7.96 (s, 1H), 7.88-7.83 (m, 2H), 7.13 (d, J=8.5 Hz, 2H), 4.26-4.11 (m, 2H), 3.77-3.46 (m, 6H), 2.78-2.65 (m, 5H), 2.48-2.34 (m, 4H), 1.76-1.36 (m, 4H).

k) (R)-4-hydroxy-7-(1-hydroxy-2-(4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)ethoxy)benzylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

HCl (2M solution in diethyl ether, 0.29 mL) was added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one acetate (Example 1, step d) (0.17 g) in EtOAc (1 mL) The resulting solution is added to 4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl) Ethoxy)benzaldehyde (Example 1, step j) (0.25 g) in EtOAc (4 mL) The reaction was cooled to 0 ° C and sodium triacetoxyborohydride (0.19 g) was added portionwise. The resulting mixture was allowed to warm to room temperature and stirred for 16 h. The reaction was partitioned between pH 7.2 phosphate buffer (50 mL) and ethyl acetate (50 mL). The layers were separated and the aqueous was extracted with ethyl acetate (2×50 mL). The aqueous phase was adjusted with sodium bicarbonate and extracted with ethyl acetate (3×50 mL). The combined organic solution was evaporated in vacuo. The residue was redissolved in EtOAc (50 mL)EtOAcEtOAc Toluene (50 mL) was added and the mixture was evaporated in vacuo. The resulting gum was dissolved in a mixture of acetonitrile and water (1:1, 10 mL) and filtered. Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 30% acetonitrile). The title compound was obtained as a white solid. The yield was 0.35 g.

m/z 640 (M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D _{6 -} DMSO, 90 ° C) δ 11.26 (s, 1H), 7.91 (s, 1H), 7.46 (d, J = 8.5 Hz, 2H), 7.03 (d, J = 8.5 Hz, 2H), 6.89 (d, J = 8.5 Hz, 1H), 6.75 (d, J = 8.2 Hz, 1H), 4.93-4.87 (m, 1H), 4.40-4.35 (m, 2H), 4.19-4.13 (m, 2H), 3.76-3.63 (m, 6H), 3.58-3.51 (m, 2H), 3.43-3.35 (m, 2H), 3.30-3.14 (m , 2H), 3.03-2.96 (m, 2H), 2.68 (s, 3H), 2.11-1.99 (m, 2H), 1.92-1.78 (m, 2H). Five unobserved exchangeable protons.

Example 2 (R)-4-hydroxy-7-(1-hydroxy-2-(9-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)decylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (9-(9-Hydroxymercapto)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-methylthiazol-4-yl)- ketone

Add 9-bromodecane-1-ol (0.29 g) to (2-methylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4 a suspension of the ketone hydrochloride (Example 1, step f) (0.4 g) in a mixture of triethylamine (0.41 mL) and acetonitrile (10 mL). The resulting mixture was stirred at 50 ° C for 16 h. The solvent was evaporated in vacuo <RTI ID=0.0> The layers were separated and the aqueous was extracted with ethyl acetate (2×30 mL). The combined organic solution was washed with brine (30 mL) dried over sodium sulfate. The residue was dissolved in MeOH (10 mL) and applied to EtOAc EtOAc. The column was washed with methanol (10 mL) and dissolved (3 mL) with 3M aqueous methanol. The eluent was evaporated in vacuo to give the subtitle compound as a yellow oil. The yield was 0.32 g.

_{^{1 H NMR (300MHz, D 6}} -DMSO) δ 7.95 (s, 1H), 4.30 (t, J = 5.1Hz, 1H), 3.78-3.44 (m, 8H), 3.42-3.33 (m, 2H), 2.69 (s, 3H), 2.35-2.14 (m, 8H), 1.71-1.57 (m, 2H), 1.55-1.19 (m, 12H).

b) 9-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)furfural

DMSO (0.32 mL) and triethylamine (0.32 mL) were added to (9-(9-hydroxyindenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4- A solution of (2-methylthiazol-4-yl)methanone (Example 2, step a) (0.32 g) in dichloromethane (5 mL). The mixture was cooled in an ice-salt bath and pyridine sulfur trioxide (0.36 g) was added. The reaction was stirred at -10 °C for 1 h, then allowed to warm to rt and stirred for additional 3 h. The reaction was diluted with DCM (20 mL) then poured into brine (20 mL). The layers were separated and EtOAc (EtOAc)EtOAc. Purification by silica gel chromatography, eluting with 47.5:47.5:5 hexanes:EtOAc:EtOAc:EtOAc: The yield is 0.25 g.

¹ H NMR (300 MHz, D ₆ -DMSO) δ 9.66 (t, J = 1.6 Hz, 1H), 7.95 (s, 1H), 3.72-3.46 (m, 8H), 3.31 (s, 2H), 2.69 ( s, 3H), 2.44 - 2.18 (m, 8H), 1.73-1.14 (m, 12H)

c) (R)-4-hydroxy-7-(1-hydroxy-2-(9-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)nonylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

( R )-7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.17g) added to 9-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)furfural (Example 2, step b) (0.23 g) and acetic acid (0.03 mL) in methanol (15 mL). The resulting mixture was stirred for 10 min and cooled to 0 °C. Sodium triethoxy borohydride (0.17 g) was then added and the mixture was stirred for 16 h. The reaction was concentrated in vacuo and the residue was crystalljjjjjjjjjj Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 30% acetonitrile). The fractions containing the product were combined and evaporated in vacuo. The residue was triturated with EtOAc (EtOAc)EtOAc. The yield was 0.15 g.

m/z 632 (M+H) ⁺ (APCI) ¹ H NMR (300 MHz, D _{6 -} DMSO, 90 ° C) δ 11.42 (s, 1H), 10.75 (s, 1H), 9.01 (s, 1H), 8.72 (s, 1H), 7.92 (s, 1H), 6.94 (d, J = 8.5 Hz, 1H), 6.78 (d, J = 8.5 Hz, 1H), 5.02-4.93 (m, 1H), 3.78-3.56 ( m,6H),3.33-3.23(m,1H),3.07-2.88(m,8H),2.69(s,3H),2.07-1.89(m,4H),1.75-1.62(m,4H),1.3- 1.24 (m, 10H) +1 protons blurred by solvent peaks.

Example 3 (R)-4-hydroxy-7-(1-hydroxy-2-(4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)ethoxy)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one dihydrochloride

a) 2-(4-(2,2-diethoxyethoxy)phenyl)ethanol

Cesium carbonate (28.3 g) was added to a solution of 4-(2-hydroxyethyl)phenol (10 g) and 2-bromo-1,1-diethoxyethane (11.79 mL) in DMF (150 mL) . The resulting suspension was heated at 90 ° C for 16 h. The reaction was poured into water (500 mL). The aqueous phase was extracted with ethyl acetate (3×200 mL). The combined organic solution was washed with water (2OmL) and brine (200 mL). Purification by gel chromatography on isohexane to 1:1 diethyl ether: EtOAc (EtOAc) The yield is 10g.

_{^{1 H NMR (300MHz, CDCl 3}} ) δ7.14 (d, J = 6.9Hz, 2H), 6.88 (d, J = 6.9Hz, 2H), 4.83 (t, J = 5.0Hz, 1H), 4.00 (d , J=5.0 Hz, 2H), 3.87-3.70 (m, 4H), 3.70-3.56 (m, 2H), 2.81 (t, J = 6.4 Hz, 2H), 1.25 (t, J = 6.9 Hz, 6H) . No OH was observed.

b) 2-(4-(2-hydroxyethyl)phenoxy)acetaldehyde

Concentrated hydrochloric acid (5 mL) was added to 2-(4-(2,2-diethoxyethoxy)phenyl)ethanol (Example 3, step a) (0.76 g) in 1,4-dioxane In a solution (10 mL), and the resulting mixture was stirred for 1 h. The reaction was diluted with water (50 mL) andEtOAcEtOAc The combined organic solution was washed with water (50 mL) EtOAc. The yield was 0.35 g.

c) (9-(2-(4-(2-hydroxyethyl)phenoxy)ethyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl (2-methylthiazol-4-yl)methanone

(2-Methylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone hydrochloride (Example 1, step f (0.63g) a mixture of 2-(4-(2-hydroxyethyl)phenoxy)acetaldehyde (Example 3, step b) (0.541g) in NMP (10mL) and acetic acid (0.11mL) In the solution. The resulting mixture was stirred at room temperature for 30 min then cooled in an ice bath. Sodium triethoxy borohydride (0.64 g) was then added and the reaction allowed to warm to rt and stirred 16 h. The reaction was diluted with methanol (30 mL) and applied to an SCX column pre-wetted with methanol. The column was washed with methanol (100 mL) and dissolved in a 3M aqueous methanol solution (100 mL). The eluent was evaporated in vacuo and the residue was purified eluting elut elut elut eluting The yield was 0.74 g.

¹ H NMR (300 MHz, D ₆ -DMSO) δ 7.86 (s, 1H), 7.09 (d, J = 8.4 Hz, 2H), 6.84-6.77 (m, 2H), 4.24 - 4.15 (m, 1H), 4.02 (t, J = 6.0 Hz, 2H), 3.68-3.54 (m, 8H), 3.00 (s, 2H), 2.71-2.61 (m, 5H), 2.51-2.42 (m, 4H), 1.75-1.65 ( m, 2H), 1.59-1.45 (m, 2H).

d) 2-(4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl) Ethoxy)phenyl)acetaldehyde

Adding trifluoroacetic acid (0.04 mL) to 9-(2-(4-(2-hydroxyethyl)phenoxy)ethyl)-4-(2-methylthiazole-4-carbonyl)-1-oxo A solution of the hetero-4-aza-9-azinospiro[5.5]undecane (Example 3, step c) (0.22 g) in DCM (3 mL) Dess-Martin periodinane (0.31 g) was then added and the resulting mixture was stirred for 5 min. Saturated sodium thiosulfate solution (0.5 mL), sodium bicarbonate solution (0.5 mL) and diethyl ether (5 mL) were then added and the mixture was stirred for 5 min. The organic layer was separated and washed with EtOAc EtOAc (EtOAc) The yield was 0.19 g.

e) (R)-4-hydroxy-7-(1-hydroxy-2-(4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)ethoxy)phenethylamino)ethyl)benzene well [d]thiazole-2(3H)-one dihydrochloride

( R )-7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.10 g) was added to 2-(4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane- A solution of 9-yl)ethoxy)phenyl)acetaldehyde (Example 3, step d) (0.14 g) and acetic acid (0.02 mL) in methanol (1 mL). Sodium triethoxy borohydride (0.103 g) was then added and the reaction was stirred 10 min then evaporated in vacuo. The residue was dissolved in a mixture of acetonitrile and water (1:1, 5 mL). Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile). The fractions containing the product were combined and evaporated in vacuo. The residue was dissolved in EtOAc (5 mL)EtOAcEtOAcEtOAc. The residue was triturated with EtOAc (EtOAc)EtOAc. The yield was 0.075 g.

m/z 654 (M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D _{6 -} DMSO) δ 11.50-11.14 (m, 2H), 9.22 (s, 1H), 8.87 (s, 1H), 7.91 ( s, 1H), 7.20 (d, J = 8.5 Hz, 2H), 6.98-6.92 (m, 3H), 6.78 (d, J = 8.5 Hz, 1H), 5.0-4.96 (m, 1H), 4.42 (t , J=5.1 Hz, 2H), 3.83-3.55 (m, 5H), 3.52-3.43 (m, 5H), 3.22-3.04 (m, 4H), 3.01-2.93 (m, 2H), 2.69 (s, 3H) ), 2.1-1.93 (m, 4H) + 2 protons blurred by solvent.

Example 4 (R)-4-hydroxy-7-(1-hydroxy-2-(2-(5-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-) Diazaspiro[5.5]undecane-9-yl)ethyl)thiophen-2-yl)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoro Acetate

a) tert-butyldimethyl(2-(thiophen-2-yl)ethoxy)decane

Third - butyldimethylsilyl group silicon chloride (12.66 g) was added portionwise As (35mL) of the 2- (2-thienyl) ethanol (9.0 g of) and imidazole (5.7 g of) in DMF. The resulting solution was stirred for 18 h. The reaction mixture was partitioned between ethyl acetate and water. The organic layer was washed with water, dried over sodium sulfate and evaporated in vacuo. Purification by gel chromatography eluting with 99:1 to 96:4 ethyl acetate:hexanes The yield is 16g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.13 (d, 1H), 6.92 (dd, J = 5.0, 3.2 Hz, 1H), 6.83-6.82 (m, 1H), 3, 82 (t, J = 6.7 Hz, 2H), 3.03 (t, J = 6.8 Hz, 2H), 0.97 (s, 9H), 0.03 (s, 6H).

b) 5-(2-(Third-butyldimethylmethylalkyloxy)ethyl)thiophene-2-carbaldehyde

N-butyllithium (in hexanes 2.5M, 30mL) was added dropwise to a third - butyldimethyl (2- (thiophen-2-yl) ethoxy) Silane (Example 4, step a) ( 16 g) in tetrahydrofuran (250 mL) in a solution at -78 °C. The reaction mixture was allowed to warm to 0 ° C and stirred for 1 h. The reaction was then cooled to -78 °C and DMF (34 mL) was added over 10 min. The reaction mixture was allowed to warm to rt and stirred for 18 h. The reaction mixture was partitioned between water and ethyl acetate. The organic layer was separated, washed with water, dried over sodium sulfate, filtered and evaporated. Purification by silica gel chromatography, eluting with EtOAc (EtOAc:EtOAc) The yield was 15.4 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ9.83 (s, 1H), 7.61 (d, J = 3.6Hz, 1H), 6.96 (d, 1H), 3.86 (t, J = 6.3Hz, 2H), 3.06 (td, J = 6.1, 0.1 Hz, 2H), 0.88 (t, J = 2.9 Hz, 9H), 0.02 (s, 6H).

c) (5-(2-(Terve-butyldimethylsilyloxy)ethyl)thiophen-2-yl)methanol

Sodium borohydride (1.74 g) was added to 5-(2-(t-butyldimethyl decyloxy)ethyl)thiophene-2-carbaldehyde (12.4 g) (Example 4, step b) Ethanol (120 mL) in a solution at 0 °C. The resulting solution was stirred at 0 ° C for 1 h. The reaction mixture was partitioned between brine and ethyl acetate. The organic layer was separated, dried over sodium sulfate, filtered and evaporated The yield was 12.1 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ6.82 (d, J = 3.6Hz, 1H), 6.69 (d, J = 3.3Hz, 1H), 4.75 (d, J = 4.9Hz, 2H), 3.81 (t , J = 6.7 Hz, 2H), 2.99 (t, J = 6.8 Hz, 2H), 1.65 (t, J = 5.5 Hz, 1H), 0.89 (d, J = 2.8 Hz, 9H), 0.03 (d, J =3.1 Hz, 6H).

d) 2-(5-(2-(Third-butyldimethylsilyloxy)ethyl)thiophen-2-yl)acetonitrile

Triphenylphosphine (13.04g) and carbon tetrabromide (15.71 g) was added to a (5- (2- (tertiary - butyl dimethyl silicone alkyloxy) ethyl) thiophen-2-yl) methanol ( Example 4, step c) (10.94 g) in DCM (20 mL) in EtOAc. The resulting solution was stirred at room temperature for 1 h. The reaction mixture was cooled to 0 ° C and tetraethylammonium cyanide (8.96 g) was added. The mixture was diluted with dichloromethane (10 mL) and stirred at rt 40 min. The reaction mixture was partitioned between dichloromethane and brine. The organic layer was separated, dried over sodium sulfate, filtered and evaporated Purification by chromatography on silica gel eluting with EtOAc (EtOAc:EtOAc) The yield was 7.6 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ6.87-6.84 (m, 1H), 6.70-6.68 (m, 1H), 3.84 (d, J = 0.8Hz, 2H), 3.80 (t, J = 6.4Hz, 2H), 2.97 (t, J = 6.5 Hz, 2H), 0.89 (s, 9H), 0.03 (s, 6H).

e) 2-(5-(2-hydroxyethyl)thiophen-2-yl)acetic acid

2-(5-(2-(Third-butyldimethylsilyloxy)ethyl)thiophen-2-yl)acetonitrile (Example 4, step d) (3 g) in ethanol (30 mL) The solution was added to a stirred solution of potassium hydroxide (1.20 g) in water (30 mL). The resulting mixture was stirred at 100 ° C for 4 hours. The reaction was concentrated in vacuo and the mixture formed was dissolved between brine and ethyl acetate. The aqueous layer was ice-cooled, acidified by dropwise addition of concentrated hydrochloric acid and extracted three times with ethyl acetate. The combined organic solution was washed with EtOAc (EtOAc m. The yield was 1.75 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ12.44 (s, 1H), 6.72 (d, J = 3.3Hz, 1H), 6.67 (d, J = 3.3Hz, 1H), 4.76 (s, 1H) , 3.71 (s, 2H), 3.61-3.55 (m, 2H), 2.86 (t, J = 6.8 Hz, 2H).

f) 2-(5-(2-(Third-butyldimethylsilyloxy)ethyl)thiophen-2-yl)ethanol

Third - butyldimethylsilyl group silicon chloride (6.63g) was added portionwise over 20 min to imidazole (2.99 g of) and 2- (5- (2-hydroxyethyl) thiophen-2-yl) acetic acid (3.9g (Example 4, step e) in a solution in DMF (50 mL). The resulting solution was stirred for 1 h. Then THF (50 mL) was added and the reaction was cooled in an ice bath. A solution of potassium carbonate (4.05 g) in water (50 mL) was then added and the mixture was stirred for 20 min. The reaction was partitioned between ethyl acetate and brine. The organic layer was separated and washed twice brine then dried over sodium sulfate. The residue was dissolved in THF (EtOAc) (EtOAc)EtOAc. The resulting solution was stirred for 2 h and the reaction was quenched (30 mL). The solvent is then evaporated in vacuo. Purification by gel chromatography eluting with 83:17 isohexane:ethyl acetate afforded the subtitle compound as a yellow liquid. The yield was 4.6 g.

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 6.69-6.63 (m, 2H), 3.87-3.75 (m, 4H), 3.05-2.91 (m, 4H), 0.89 (s, 9H), 0.03 (s, 6H) + exchangeable protons

g) 2-(5-(2-(Third-butyldimethylsilyloxy)ethyl)thiophen-2-yl)acetaldehyde

Dess - Martin periodinane (0.38 g) was added to 2- (5- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethyl) thiophen-2-yl) ethanol (Example 4 , step f) (0.22 g) in dichloromethane (5 mL) A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (25 mL). The organic layer was separated and washed with EtOAc EtOAc m. The yield was 0.21 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 9.70 (t, J = 2.2 Hz, 1H), 6.73 (s, 2H), 3.83 - 3.78 (m, 4H), 2.98 (t, J = 6.7 Hz, 2H), 0.89 (s, 9H), 0.02 (s, 6H)

h) (2-methylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate

1-Propanephosphonic acid cyclic anhydride (1.57 M solution in THF, 2.49 mL) was added to 1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester salt A solution of the acid salt (WuXi PharmaTech) (1 g), 2-methylthiazole-4-carboxylic acid (0.56 g) and triethylamine (3.26 mL) in DMF (30 mL) . The reaction was partitioned between water (500 mL) and ethyl acetate (200 mL). The layers were separated and the aqueous layer was extracted with ethyl acetate (2×150 mL). The combined organic solution was washed with water (2×100 mL) and brine (100 mL). Purify by gel chromatography and elute with ethyl acetate. The oil formed was dissolved in dichloromethane (30 mL) and trifluoroacetic acid (3 mL) was added dropwise. This was stirred for 1 hour and concentrated in vacuo. The residue was azeotroped twice with toluene (20 mL). The resulting gum was triturated with diethyl ether to give the subtitle compound as a white solid. The yield was 1.20 g.

m/z 282 (M+H) ⁺ (APCI) ¹ H NMR (300MHz, D6-DMSO) δ 8.59-8.18 (m, 2H), 8.00 (s, 1H), 3.86-3.49 (m, 6H), 3.22 -2.86 (m, 4H), 2.69 (s, 3H), 2.00-1.90 (m, 2H), 1.74-1.58 (m, 2H).

i) (9-(2-(5-(2-(Third-butyldimethylsilyloxy)ethyl)thiophen-2-yl)ethyl)-1-oxa-4,9- Diazaspiro[5.5]undecane-4-yl)(2-methylthiazol-4-yl)methanone

(2-Methylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 4, steps h) (0.25 g) was added to 2-(5-(2-(t-butyldimethyl decyloxy)ethyl)thiophen-2-yl)acetaldehyde (0.2 g) (Example 4, Step g) is a solution in a mixture of NMP (5 mL) and acetic acid (0.04 mL). The resulting mixture was stirred for 5 min then sodium triethoxysulfonylborohydride (0.22 g) was added. The mixture was stirred for 1 h and poured into a pH 7.2 buffer (50 mL). The aqueous phase was extracted with ethyl acetate (3×50 mL). The combined organics were washed with aq. EtOAc (50 mL) and brine (50 mL). Purification by silica gel chromatography eluting with 4:1:0.05HHHHHHHHHHHHHHHHHH The yield was 0.23 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 7.86 (s, 1H), 6.62 (s, 2H), 3.76 (t, J = 6.5 Hz, 2H), 3.70 - 3.57 (m, 6H), 3.30 ( t, J=7.0HZ, 2H), 2.91-2.81 (m, 4H), 2.68 (s, 3H), 2.44-2.37 (m, 4H), 1.74-1.66 (m, 2H), 1.58-1.49 (m, 2H), 0.88-0.84 (m, 9H), 0.01-0.00 (m, 6H).

j) (9-(2-(5-(2-hydroxyethyl)thiophen-2-yl)ethyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4 -yl)(2-methylthiazol-4-yl)methanone

Concentrated hydrochloric acid (0.5 mL) was added to (9-(2-(5-(2-(t-butyl dimethyl decyloxy)ethyl) thiophen-2-yl)ethyl)-1- Oxa-4,9-diazaspiro[5.5]undecethane-4-yl)(2-methylthiazol-4-yl)methanone (Example 4, step i) (0.235 g) in methanol The solution in (5 mL) and the resulting solution was stirred for 1 h. The solvent was evaporated in vacuo and the residue was taken to toluene toluene and then dissolved in methanol (~2mL). The residue was dissolved in MeOH (10 mL) and applied to EtOAc EtOAc. The column was washed with methanol (100 mL) and dissolved in 1M aqueous methanol (100 mL). The eluent was evaporated in vacuo to give the subtitle compound as a yellow oil. The yield was 0.12 g.

_{^{1 H NMR (300MHz, D 6}} -DMSO) δ7.87 (s, 1H), 6.64 (s, 2H), 4.38 (t, J = 4.9Hz, 1H), 3.70-3.51 (m, 8H), 2.84 ( t, J = 6.7 Hz, 4H), 2.68 (s, 3H), 2.60-2.52 (m, 2H), 2.4-2.38 (m, 4H), 1.76-1.65 (m, 2H), 1.60-1.49 (m, 2H).

k) (R)-4-hydroxy-7-(1-hydroxy-2-(2-(5-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4), 9-diazaspiro[5.5]undecane-9-yl)ethyl)thiophen-2-yl)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one di- Trifluoroacetate

Adding trifluoroacetic acid (0.02 mL) to (9-(2-(5-(2-hydroxyethyl)thiophen-2-yl)ethyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-4-yl)(2-methylthiazol-4-yl)methanone (Example 4, step j) (0.128 g) in DCM (5 mL) in EtOAc The mixture was stirred for 5 min. Dess-Martin periodinane (0.15 g) was then added and the mixture was stirred for 10 min. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (25 mL). The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with EtOAc EtOAc (EtOAc) The residue was redissolved in MeOH (1 mL). Acetic acid (0.04 mL) was added followed by ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.19 g) and the mixture was cooled to 0 °C. Sodium triethoxysulfonium borohydride (0.157 g) was added and the resulting mixture was stirred 1 h then concentrated in vacuo. The residue was dissolved in a mixture of acetonitrile and water (1:1, 5 mL). Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile). The title compound was obtained as a white solid. The yield was 0.04 g.

m/z 644 (M+H) ⁺ (APCI) ¹ H NMR (300 MHz, D6-DMSO, 90 ° C) δ 7.92 (s, 1H), 6.93 (d, J = 8.5 Hz, 1H), 6.84-6.73 (m,3H), 4.89 (dd, J=7.9, 5.4 Hz, 1H), 3.74-3.62 (m, 6H), 3.44-3.07 (m, 14H), 2.68 (s, 3H), 2.11-1.93 (m , 2H), 1.89-1.72 (m, 2H). Six unobserved exchangeable protons.

Example 5 (R)-4-hydroxy-7-(1-hydroxy-2-(4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)ethyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2,2'-(1,4-phenylene)diethanol

Add borane-methylthio complex (2M solution in tetrahydrofuran, 80 mL) to 2,2'-(1,4-phenylene)diacetic acid (10.20 g) in tetrahydrofuran (100 mL) over 20 min Cool the stirred solution in an ice bath. After 16 h, the reaction mixture was cautiously quenched with methanol (40 mL). The solution was evaporated in vacuo and the formed gum was dissolved between ethyl acetate and saturated aqueous ammonium chloride. The ethyl acetate solution was separated, dried over magnesium sulfate, filtered and evaporated in vacuo The yield was 8.64 g.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.18 (s, 4H), 3.86 (t, 4H), 2.85 (t, 4H) + 2 exchangeable protons.

b) 2-{4-[2-(Third-butyl-dimethyl-decyloxy)-ethyl]-phenyl}-ethanol

Third - Silane-butyldimethylsilyl chloride (9.68mL) was added to 2,2 '- (1,4-phenylene) diethanol (example 5, step a) (8.64g) and imidazole (10.21 g) It was cooled in an ice bath in anhydrous DMF (100 mL). After 45 min the reaction mixture was diluted with EtOAc EtOAc. Purification by gel chromatography eluting with 5:1 isohexane:ethyl acetate afforded the subtitle compound as colorless oil. The yield was 6.20 g.

¹ H NMR (300MHz, CDCl ₃ ) δ 7.21 - 7.13 (m, 4H), 3.92-3.76 (m, 4H), 2.92-2.76 (m, 4H), 0.89 (s, 9H), 0.02 (s, 6H) )

c) 2-(4-(2-(Tern-butyldimethyl)alkyl)ethyl)phenyl)acetaldehyde

Dess - Martin periodinane (0.38 g) was added to 2- (4- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethyl) phenyl) ethanol (example 5, step b (0.21 g) in a solution of dichloromethane (5 mL) and the mixture was stirred for 30 min. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (25 mL). The organic phase was separated and washed with EtOAc EtOAc EtOAc (EtOAc) The yield was 0.20 g.

¹ H NMR (300 MHz, CDCl ₃ ) δ 9.74 (t, J = 2.4 Hz, 1H), 7.23 (d, J = 8.2 Hz, 2H), 7.15 (d, J = 8.2 Hz, 2H), 3.82 (t , J = 7.1 Hz, 2H), 3.67 (d, J = 2.6 Hz, 2H), 2.83 (t, J = 7.1 Hz, 2H), 0.88 (s, 9H), 0.00 (s, 6H).

d) (9-(4-(2-(Third-butyldimethylsilyloxy)ethyl)phenethyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkan-4-yl)(2-methylthiazol-4-yl)methanone

(2-Methylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 4, steps h) (0.26g) was added to 2- (4- (2- (third - butyldimethylsilyl silicon alkyloxy) ethyl) phenyl) acetaldehyde (example 5, step c) (0.2g) And a solution of acetic acid (0.04 mL) in NMP (5 mL). The resulting mixture was stirred for 5 min then sodium triacetoxyborohydride (0.23 g) was added. The mixture was stirred for 1 h, poured into EtOAc EtOAc (EtOAc) The combined organic solution was washed with aq. EtOAc (50 mL) and brine (50 mL). Purification by chromatography on silica gel eluting with EtOAc (EtOAc:EtOAc) The yield was 0.30 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ7.90 (s, 1H), 7.12 (s, 4H), 3.80 (t, J = 6.8Hz, 2H), 3.71-3.60 (m, 6H) , 2.77-2.67 (m, 7H), 2.46-2.41 (m, 2H), 1.76-1.67 (m, 2H), 1.61-1.51 (m, 2H), 0.87 (s, 9H), 0.02 (s, 6H) +4 protons that are obscured by solvent peaks.

e) (9-(4-(2-hydroxyethyl)phenethyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-methyl Thiazol-4-yl)methanone

Concentrated hydrochloric acid (0.5mL) was added to (9- (4- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethyl) phenethyl) -l-oxa-4,9- Azaspiro[5.5]undecane-4-yl)(2-methylthiazol-4-yl)methanone (Example 5, step d) (0.3 g) in MeOH (5 mL) The resulting solution was stirred for 1 h. The solvent was evaporated in vacuo and the residue was taken from toluene toluene and then dissolved in methanol (~2mL). The residue was diluted with methanol (10 mL) and applied to an SCX column pre-wetted with methanol. The column was washed with methanol (100 mL) and dissolved in 1M aqueous methanol (100 mL). The eluent is evaporated in vacuo to give the title compound as a clear oil. The yield was 0.2 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 7.86 (s, 1H), 7.09 (s, 4H), 4.24 (t, J = 5.0 Hz, 1H), 3.69-3.55 (m, 8H), 2.68 ( s, 3H), 2.45-2.38 (m, 4H), 1.74-1.65 (m, 2H), 1.57-1.48 (m, 2H) + 6 protons obscured by solvent peaks.

f) (R)-4-hydroxy-7-(1-hydroxy-2-(4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)ethyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

Adding trifluoroacetic acid (0.04 mL) to (9-(4-(2-hydroxyethyl)phenethyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4 (Based) (2-methylthiazol-4-yl)methanone (Example 5, step e) (0.21 g) in DCM (5 mL) in EtOAc. Dess-Martin periodinane (0.25 g) was then added and the mixture was stirred for 10 min. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (25 mL). The aqueous solution was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with EtOAc EtOAc (EtOAc) The residue was redissolved in MeOH (1 mL). Acetic acid (0.04 mL) was added followed by ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.19 g) and the mixture was cooled to 0 °C. Sodium triethoxysulfonium borohydride (0.157 g) was added and the resulting mixture was stirred 1 h then concentrated in vacuo. The residue was dissolved in a mixture of acetonitrile and water (1:1, 5 mL). Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile). The title compound was obtained as a white solid. The yield was 0.17 g.

m/z 638 (M+H) ⁺ (APCI) ¹ H NMR (300 MHz, D _{6 -} DMSO) δ 7.92 (s, 1H), 7.26-7.18 (m, 4H), 6.93 (d, J = 8.5 Hz, 1H), 6.77 (d, J = 8.5 Hz, 1H), 4.96 - 4.87 (m, 1H), 3.76 - 2.90 (m, 20H), 2.68 (s, 3H), 2.14-1.75 (m, 4H). Six unobserved exchangeable protons.

Example 6 (R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(3-(bromomethyl)phenyl)ethanol

Borane dimethyl sulfide complex (2M solution in THF, 5.78 mL) was added dropwise to 2-(3-(bromomethyl)phenyl)acetic acid (1.06 g) in THF (10 mL) at 0 ° C The solution was stirred and the resulting mixture was stirred for 10 min. The reaction was then allowed to warm to room temperature and stirred overnight. Methanol (5 mL) was then added and the mixture was concentrated in vacuo. Purification by chromatography on silica gel eluting with EtOAc (EtOAc) The yield is 1g.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.34 - 7.24 (m, 3H), 7.22 - 7.12 (m, 1H), 4.48 (s, 2H), 3.87 (t, J = 6.5 Hz, 2H), 2.87 ( t, J = 6.5 Hz, 2H), no OH was observed.

b) 9-(3-(2-Hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-methylthiazole- 4-yl)methanone

2-(3-(Bromomethyl)phenyl)ethanol (Example 6, step a) (0.11 g) was added to (2-methylthiazol-4-yl)(1-oxa-4,9- A solution of diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 4, step h) (0.2 g) and triethylamine (0.17 mL) in acetonitrile (5 mL) The mixture formed and stirred at room temperature overnight. The solvent was evaporated in vacuo <RTI ID=0.0> The aqueous layer was separated and extracted with ethyl acetate (2×25 mL). The combined organic solution was washed with brine (25 mL) dried over sodium sulfate. Purification by silica gel chromatography, eluting with EtOAc EtOAc:EtOAc:EtOAc The yield was 0.15 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ7.85 (s, 1H), 7.18 (t, J = 7.4Hz, 1H), 7.12-7.04 (m, 3H), 4.24 (t, J = 5.1Hz, 1H), 3.71-3.54 (m, 8H), 3.42 (s, 2H), 2.71 (t, J = 6.9 Hz, 2H), 2.68 (s, 3H), 2.38-2.27 (m, 4H), 1.74-1.64 (m, 2H), 1.57-1.47 (m, 2H).

c) (R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

Adding trifluoroacetic acid (0.03 mL) to (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4- (2,4-Methylthiazol-4-yl)methanone (Example 6, step b) (0.17 g) in DCM (5 mL) in EtOAc. Dess-Martin periodinane (0.21 g) was then added and the mixture was stirred for 10 min. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (25 mL). The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with aq. sodium hydrogen sulfate (10 mL) and brine The residue was redissolved in methanol (25 mL). Acetic acid (0.04 mL) was added followed by ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.16 g) and the mixture was cooled to 0 ° C in an ice bath, then sodium triacetoxyborohydride (0.16 g). The resulting mixture was stirred for 1 h and concentrated. The residue was dissolved in a mixture of acetonitrile and water (1:1, 5 mL). Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile). The title compound was obtained as a white solid. The yield was 0.125 g.

m/z 624 (M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D _{6 -} DMSO, 90 ° C) δ 11.27 (s, 1H), 7.90 (s, 1H), 7.45-7.31 (m, 4H) ), 6.93 (d, J = 8.2 Hz, 1H), 6.77 (d, J = 8.2 Hz, 1H), 4.92 (dd, J = 7.9, 5.1 Hz, 1H), 4.30 (s, 2H), 3.74 - 3.58 (m, 6H), 3.29-2.97 (m, 10H), 2.67 (s, 3H), 2.13-1.95 (m, 2H), 1.88-1.70 (m, 2H). Five unobserved exchangeable protons.

Example 7 (R)-8-hydroxy-5-(1-hydroxy-2-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

Adding trifluoroacetic acid (0.02 mL) to (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4- (2,4-Methylthiazol-4-yl)methanone (Example 6, step b) (0.11 g) in DCM (5 mL) in EtOAc. Dess-Martin periodinane (0.14 g) was then added and the mixture was stirred for 10 min. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (25 mL). The aqueous layer was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with EtOAc EtOAc m. The residue was redissolved in methanol (25 mL). Add acetic acid (0.02 mL) followed by ( R )-5-(2-amino-1-(tris-butyldimethylsilyloxy)ethyl)-8-hydroxyquinoline-2 (1H) - Ketone (WO 2004106333) (0.09 g) and the mixture was cooled to 0 ° C in an ice-bath, then sodium triacetoxyborohydride (0.08 g). The resulting mixture was stirred for 1 h and concentrated in vacuo. The residue was partitioned between ethyl acetate (50 mL) and EtOAc (EtOAc) The aqueous solution was separated and extracted with ethyl acetate (2×50 mL). The combined organic solution was washed with brine (20 mL) dried over sodium sulfate. Purification by gel chromatography, eluting with a gradient of 95:5:0.5 to 92:8:0.8 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined, evaporated and dissolved in THF (1 mL). Triethylamine trihydrofluoride (0.05 mL) was added and the mixture was stirred overnight. The solvent was evaporated in vacuo and the residue was crystalljjjjjjjjj Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile). The title compound was obtained as a white solid. The yield was 0.06 g.

m/z 618 (M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D _{6 -} DMSO) δ 8.16 (d, J = 10.0 Hz, 1H), 7.90 (s, 1H), 7.47-7.30 (m) , 4H), 7.13 (d, J = 8.2 Hz, 1H), 6.99 (d, J = 8.2 Hz, 1H), 6.54 (d, J = 10.0 Hz, 1H), 5.35 (dd, J = 8.5, 4.1 Hz) ,1H),4.30(s,2H),3.74-3.57(m,4H),3.32-2.98(m,12H),2.67(s,3H),2.12-1.92(m,2H),1.87-1.67(m , 2H). Six unobserved exchangeable protons.

Example 8 (R)-4-hydroxy-7-(1-hydroxy-2-(4-(3-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)propoxy)benzylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 4-(3-bromopropoxy)benzaldehyde

1,3-Dibromopropane (20.0 g) was added to a stirred suspension of 4-hydroxybenzaldehyde (4.0 g) and potassium carbonate (7.0 g) in acetone (80 mL). After 16 h, the mixture was heated at reflux for a further 2 h, cooled and filtered to remove organics and the solution evaporated in vacuo. Purification by gel chromatography, dissolving in isohexane to remove 1,3-dibromopropane, and then dichloromethane: isohexane 2:1 to collect sub-title compound. The yield was 3.45 g.

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 9.89 (s, 1H), 7.84 (d, J = 9.2Hz, 2H), 7.01 (d, J = 9.2Hz, 2H), 4.21 (t, J = 5.6Hz, 2H), 3.62 (t, J = 6.0 Hz, 2H), 2.36 (five peaks, J = 6.1 Hz, 2H)

b) 4-(3-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)propoxy Benzaldehyde

(2-methylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 4, step h (0.244g), 4-(3-bromopropoxy)benzaldehyde (Example 8, Step a) (0.15g) and a mixture of triethylamine (0.344mL) in MeCN (2mL) heated at 60 ° C . After 16 h, the reaction mixture was evaporated in vacuo. Purification by silica gel chromatography, eluting with ethyl acetate: triethylamine 10:1 to give the subtitle compound. The yield was 0.24 g.

m/z 444(M+H) ⁺ (APCI)

c) (R)-4-hydroxy-7-(1-hydroxy-2-(4-(3-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)propoxy)benzylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

Hydrogen chloride (0.4 mL of 1 M diethyl ether solution) was added to 4-(3-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)propoxy)benzaldehyde (Example 8, Step b) (0.15 g) and ( R )-7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzene And [d] a stirred solution of thiazole-2(3H)-one acetate (Example 1, step d) (0.15 g) in MeOH (15 mL). After 5 minutes, sodium cyanoborohydride (0.05 g) was added. After 16 h, the solution was concentrated to ~2 mL and then partitioned between ethyl acetate and pH 7.2 phosphate buffer (20 mL). The ethyl acetate solution was washed with brine, dried over sodium sulfate, filtered and evaporated in vacuo. The residue was dissolved in a mixture of acetonitrile and water (1:1, 5 mL). Purification by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 5-35% acetonitrile). The title compound was obtained as a white solid. The yield was 0.045 g.

m/z 654 (M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D _{6 -} DMSO) δ 7.92 (s, 1H), 7.42 (d, J = 8.7 Hz, 2H), 6.98 (d, J) =8.7 Hz, 2H), 6.89 (d, J = 8.5 Hz, 1H), 6.75 (d, J = 8.2 Hz, 1H), 4.91-4.84 (m, 1H), 4.18-4.04 (m, 4H), 3.78 - 2.94 (m, 14H), 2.71 (s, 3H), 2.19 - 2.00 (m, 4H), 1.85-1.66 (m, 2H). Six unobserved exchangeable protons.

Example 9 (R)-8-hydroxy-5-(1-hydroxy-2-(3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

a) 4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester

1-Propanephosphonic acid cyclic anhydride (1.57 M solution in THF, 0.64 mL) was added to 1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester salt A solution of the acid salt (WuXi PharmaTech) (0.26 g), 5-methylthiophene-2-carboxylic acid (0.142 g) and triethylamine (0.84 mL) in DMF (8 mL). The reaction mixture was poured into water (100 mL) andEtOAcEtOAc The combined organic solution was washed with water (2×100 mL) and brine (100 mL). Purification by chromatography on silica gel eluting with EtOAc (EtOAc) The yield was 0.32 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.11 (d, J = 3.6Hz, 1H), 6.72-6.69 (m, 1H), 3.78-3.67 (m, 8H), 3.60-3.51 (m, 2H), 3.19-3.10 (m, 2H), 2.51 (s, 3H), 1.86-1.79 (m, 2H), 1.45 (s, 9H).

b) (5-methylthien-2-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate

3-(5-Methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester (Example 9, step a) (0.32 g) was treated with trifluoroacetic acid (1.0 g) in DCM (3 mL). After 2 h, the reaction mixture was evaporated in vacuo and aq. The yield was 0.32 g.

c) (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(5-methylthiophene) -2-yl) ketone

2-(3-(Bromomethyl)phenyl)ethanol (Example 6, step a) (0.136 g) was added to (5-methylthiophen-2-yl)(1-oxa-4,9- Stirring of diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 9, step b) (0.250 g) and triethylamine (0.278 mL) in MeCN (5 mL) In solution. After 1 h, the reaction mixture was concentrated and applied to a silica gel column eluting with ethyl acetate: triethylamine 95:5 to give sub-title compound. The yield was 0.24 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.28-7.24 (m, 2H), 7.18-7.15 (m, 1H), 7.11-7.08 (m, 2H), 6.70-6.67 (m, 1H), 3.86 (t , J=6.6 Hz, 2H), 3.77-3.69 (m, 4H), 3.56 (s, 2H), 3.48 (s, 2H), 2.86 (t, J = 7.0 Hz, 2H), 2.55-2.48 (m, 5H), 2.40-2.32 (m, 2H), 1.89-1.82 (m, 2H), 1.70-1.50 (2H at the water peak). An unobserved exchangeable proton.

d) 2-(3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl Phenyl) acetaldehyde

Dess-Martin periodinane (0.16 g) was added to (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] under nitrogen. Stupidization of undecane-4-yl)(5-methylthiophen-2-yl)methanone (0.13 g) (Example 9, step c) and trifluoroacetic acid (0.036 g) in DCM (4 mL) In solution. After 0.5 h, a saturated aqueous solution of sodium thiosulfate (5 mL) and a saturated sodium hydrogen carbonate solution (5 mL) was added and the mixture was extracted twice with ethyl acetate. The combined organic extracts were washed with EtOAc EtOAc m. The yield was 0.13 g. Use directly.

e) (R)-8-hydroxy-5-(1-hydroxy-2-(3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

The (R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8-hydroxy-quinolin -2 (1H) - one (WO 2004106333) ( 0.179 g) added to 2-(3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl) a stirred solution of methyl)phenyl)acetaldehyde (Example 9, step d) (0.13 g) and acetic acid (0.031 mL) in MeOH (5 mL). After 5 minutes, sodium triethoxysulfonium borohydride (0.114 g) was added. After a further 15 min, ethyl acetate (30 mL) was added and the mixture was washed with &lt Purification by gel chromatography, eluting with 92:7:1, DCM:MeOH: ' This intermediate was dissolved in THF (2 mL) and triethylamine <RTI ID=0.0> After 16 h the reaction was evaporated in vacuo <~~~~~~ The residue was dissolved in a mixture of acetonitrile and water (1:1, 5 mL). Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile). The title compound was obtained as a white solid. The yield was 0.05 g.

m/z 617 (M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D _{6 -} DMSO) δ 10.55-10.45 (m, 2H), 9.95-9.84 (m, 0.2H), 9.68-9.57 (m , 0.8H) (rotational isomer), 8.97-8.74 (m, 2H), 8.16 (d, J = 9.5 Hz, 1H), 7.49-7.33 (m, 4H), 7.29-7.26 (m, 0.2H) , 7.25-7.21 (m, 0.8H)) (rotational isomer), 7.15 (d, J = 9.5 Hz, 1H), 6.99 (d, J = 9.5 Hz, 1H), 6.86-6.81 (m, 1H) , 6.58 (d, J = 10.1 Hz, 1H), 6.27-6.18 (m, 1H), 5.37-5.30 (m, 2H), 4.44 - 4.40 (m, 0.5H), 4.35-4.29 (m, 1.5H) , 3.73-3.63 (m, 4H), 3.51-3.46 (m, 2H), 3.31-2.94 (m, 8H), 2.46 (s, 3H), 2.12-2.00 (m, 2H), 1.70-1.58 (m, 2H). An unobserved exchangeable proton.

Example 10 (R)-4-hydroxy-7-(1-hydroxy-2-(3-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)ethoxy)benzylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(3-methylnonylphenoxy)ethyl methanesulfonate

2-Bromoethanol (10.23 g) and potassium carbonate (11.32 g) were added to a solution of 3-hydroxybenzaldehyde (5 g) in acetonitrile (100 mL). The reaction mixture was cooled and partitioned between EtOAc and EtOAc. The organic layer was separated, washed with brine, dried over sodium sulfate, filtered and evaporated. The residue was dissolved in DCM (30 mL)EtOAc. The solution was cooled to 0 ° C and treated with methane sulfonium chloride (1.89 mL). The reaction mixture was stirred at 0 ° C for 10 min and then allowed to warm to rt and stirred 1 h. The mixture was washed twice with brine, dried over sodium sulfate, filtered and evaporated. Purified by silica gel chromatography, eluting with 6:4 ethyl acetate: isohexane. The pure fractions were evaporated in vacuo to give the subtitle compound as a colourless oil. The yield was 3.50 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ9.98 (d, J = 8.2Hz, 1H), 7.53-7.46 (m, 2H), 7.41-7.39 (m, 1H), 7.22-7.19 (m, 1H), 4.62-4.59 (m, 2H), 4.34-4.30 (m, 2H), 3.10 (s, 3H).

b) 3-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethoxylate Benzaldehyde

2-(3-Mercaptophenoxy)ethyl methanesulfonate (Example 10, step a) (0.38 g) was added to (2-methylthiazol-4-yl)(1-oxo-4) ,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 4, step h) (0.62 g) and triethylamine (0.55 mL) in acetonitrile (5 mL) The solution was stirred and the mixture formed was stirred at 65 ° C for 16 h. The solvent was evaporated in vacuo <RTI ID=0.0> The aqueous phase was separated and extracted with ethyl acetate (2×25 mL). The organic phase was combined, washed with brine (25 mLs Purification by silica gel chromatography, eluting with 47.5:47.5:5 hexanes:EtOAc:EtOAc:EtOAc: The yield was 0.48 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ.9.97 (s, 1H), 7.96 (s, 1H), 7.56-7.40 (m, 3H), 7.32-7.25 (m, 1H), 4.14 (s, 2H ), 3.75-3.47 (m, 6H), 2.80-2.62 (m, 5H), 2.55-2.37 (m, 4H), 1.74-1.40 (m, 4H).

c) (R)-4-hydroxy-7-(1-hydroxy-2-(3-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)ethoxy)benzylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

3-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethoxy) Benzaldehyde (Example 10, step b) (0.24 g) was added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)- A mixture of ketone hydrochloride (WO 2007027134, Example 1, step d) (0.15 g) and acetic acid (0.032 mL) in methanol (2 mL). The mixture was stirred for 30 min and then cooled in an ice bath. Sodium triethoxy borohydride (0.18 g) was then added and the mixture was stirred for 2 h. The solvent was evaporated in vacuo and EtOAcqqqqqqqqqq The aqueous phase was separated and extracted with ethyl acetate (2×50 mL). The combined organic phases were washed with brine (20 mL) dried over sodium sulfate. Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile). The title compound was obtained as a white solid. The yield was 0.21 g.

m/z 640 (M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D _{6 -} DMSO, 90 ° C) δ 11.27 (s, 1H), 7.92 (s, 1H), 7.37 (t, J = 7.8 Hz, 1H), 7.19-7.12 (m, 2H), 7.05 (d, J = 8.2 Hz, 1H), 6.90 (d, J = 8.2 Hz, 1H), 6.76 (d, J = 8.2 Hz, 1H), 4.92 (t, J = 6.5 Hz, 1H), 4.45 - 4.31 (m, 2H), 4.26 - 4.12 (m, 2H), 3.76 - 3.53 (m, 8H), 3.45 - 3.36 (m, 2H), 3.31 3.20 (m, 2H), 3.06-3.00 (m, 2H), 2.68 (s, 3H), 2.1-1.97 (m, 2H), 1.92-1.80 (m, 2H). Five unobserved exchangeable protons.

Example 11 5-hydroxy-8-(1-hydroxy-2-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] eleven Cycloalkane-9-yl)methyl)phenethylamino)ethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one di-trifluoroacetate, isomer Object 1

a) 1-(2,4-dihydroxy-3-nitrophenyl)ethanone

2-Nitrobenzene-1,3-diol (24.5 g) was added in portions over 15 minutes to a vigorously stirred solution of aluminum chloride (46.3 g) in nitrobenzene (325 mL). Acetic anhydride (15.7 mL) was then added dropwise to the mixture over a further 15 minutes and the mixture was then heated at 100 °C for 5 h. The reaction was cooled to ambient temperature and cautiously quenched with ice cold 2M hydrochloric acid (300 mL). The mixture was extracted with diethyl ether (2×500 mL) and combined diethyl ether extracts (2×400 mL). The combined basic extracts were washed with diethyl ether (4×500 mL) and then acidified to pH 1 with 2M hydrochloric acid (700 mL). The precipitate which formed was filtered off, washed with water and dried <RTI ID=0.0> The yield was 29.5 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ13.32 (s, 1H), 12.31 (s, 1H), 7.98 (d, J = 9.2Hz, 1H), 6.63 (d, J = 28.2Hz, 1H) , 2.59 (s, 3H).

b) 1-(4-(Benzyloxy)-2-hydroxy-3-nitrophenyl)ethanone

Lithium third butanol (4.06 g) was added to 1-(2,4-dihydroxy-3-nitrophenyl)ethanone under nitrogen at an internal temperature of less than 30 ° C (Example 11, Step a) (10 g) in a stirred solution in DMF (100 mL). After stirring for an additional 10 minutes at ambient temperature, benzyl bromide (6.03 mL) was added and the mixture was stirred for additional 20 h. Additional benzyl bromide (3 mL) was added and the mixture was stirred for 24 h. The reaction was quenched with water (300 mL) EtOAc (EtOAc)EtOAc. The basic solution was cooled in ice/water, EtOAc (EtOAc)EtOAc. The solid was triturated with EtOAc (EtOAc)EtOAc. The yield was 6.8 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ13.04 (s, 1H), 8.14 (d, J = 9.2Hz, 1H), 7.45-7.32 (m, 5H), 7.01 (d, J = 9.2Hz, 1H), 5.42 (s, 2H), 2.64 (s, 3H) + 3 unobservable exchangeable protons.

c) 1-(3-Amino-4-(benzyloxy)-2-hydroxyphenyl)ethanone

Zinc powder (5.5 g) was added in portions to 1-(4-(benzyloxy)-2-hydroxy-3-nitrophenyl group in 15 minutes while keeping the internal temperature below 40 ° C in an ice bath. Ethyl ketone (Example 11, step b) (5.5 g) in a suspension in acetic acid (55 mL). The mixture was allowed to reach ambient temperature and stirred for 2 h. The mixture was filtered through Celite (caution: heat - do not dry), washed with acetic acid and poured onto ice/water (500 mL). The resulting precipitate was filtered, washed with water and dried then dried then evaporated The yield was 4.8 g.

_{^{1 H NMR (300MHz, D 6}} -DMSO) δ7.53 (m, 2H), 7.48-7.33 (m, 3H), 7.28 (d, J = 9.0Hz, 1H), 6.72 (d, J = 9.0Hz, 1H), 5.29 (s, 2H), 2.59 (s, 3H).

d) 8-Ethyl-5-(benzyloxy)-2H-benzo[b][1,4]oxazin-3(4H)-one

2-Chloroacetyl chloride (1.8 mL) was added dropwise to 1-(3-amino-4-(benzyloxy)-2-hydroxyphenyl)ethanone (Example 11, step c) (5.2 g) and a stirred mixture of sodium bicarbonate (3.74 g) in DMF (30 mL). Cesium carbonate (7.90 g) was added and the mixture was heated at 100 °C for 20 h. The mixture was cooled to rt. EtOAc (EtOAc)EtOAc. The solid residue was taken up in EtOAc (EtOAc m. The yield was 5.7 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ 10.33 (s, 1H), 7.55 (m, 2H), 7.39 (m, 2H), 7.34 (d, J = 8.8Hz, 1H), 7.33 (m, 1H ), 6.89 (d, J = 9.2 Hz, 1H), 5.27 (s, 2H), 4.67 (s, 2H), 3.32 (s, 3H).

e) 5-(Benzyloxy)-8-(2-chloroethyl)-2H-benzo[b][1,4]oxazin-3(4H)-one

Addition of benzyltrimethylammonium dibromide (14.17 g) to 8-ethylindolyl-5-(benzyloxy)-2H-benzo[b][1,4]oxazine-3 (4H The ketone (Example 11, step d) (5.5 g) was stirred in EtOAc (EtOAc) (EtOAc) The reaction was cooled to ambient temperature and treated with aqueous sodium bisulfite (5.78 g in 100 mL) and stirred for 30 min. The mixture was diluted with EtOAc (EtOAc)EtOAc. The yield was 5.6 g.

_{^{1 H NMR (300MHz, D 6}} -DMSO) δ10.41 (s, 1H), 7.55 (m, 2H), 7.44 (d, J = 9.4Hz, 1H), 7.39 (m, 2H), 7.32 (m, 1H), 6.95 (d, J = 9.4 Hz, 1H), 5.30 (s, 2H), 4.96 (s, 2H), 4.69 (s, 2H).

f) 8-(2-azidoethyl)-5-(benzyloxy)-2H-benzo[b][1,4]oxazin-3(4H)-one

Add sodium azide (1.176 g) to 5-(benzyloxy)-8-(2-chloroethenyl)-2H-benzo[b][1,4]oxazine-3 (4H) a ketone (Example 11, step e) (4.8 g) in EtOAc m. The mixture was poured onto ice/water and the solid formed was collected by filtration, washed with water and dried <RTIgt; The yield was 4.6 g.

_{^{1 H NMR (300MHz, D 6}} -DMSO) δ10.42 (s, 1H), 7.55 (m, 2H), 7.48 (m, 1H), 7.43-7.29 (m, 3H), 6.97 (m, 1H), 5.31 (s, 2H), 4.69 (s, 2H), 4.63 (s, 2H).

g) 8-(2-azido-1-hydroxyethyl)-5-(benzyloxy)-2H-benzo[b][1,4]oxazin-3(4H)-one, different Structure 1

8-(2-azidoethyl)-5-(benzyloxy)-2H-benzo[b][1,4]oxazin-3(4H)-one (Example 11, step f The suspension of (2 g) in ethanol (80 mL) was taken with sodium borohydride (0.224 g) and the mixture was stirred at <RTIgt; The mixture was partitioned between EtOAc and EtOAcq. The crude solid was triturated with acetone (20 mL) to yield 1.6 g.

Separation of the racemic mixture:

Racemic 8-(2-azido-1-hydroxyethyl)-5-(benzyloxy)-2H-benzo[b][1,4]oxazin-3(4H)-one (2.5 g) Dissolved in methanol at a concentration of 10 mg/mL, using Chiralpak by palmitic HPLC AS (250 x 50 mm ID, 20 μm particle size) column was separated by ethanol elution. The fractions containing the first dissolved isomer are combined and concentrated in vacuo to give the sub-title compound. The yield was 0.67 g.

m/z 339 (MH) ^- (APCI)

h) 8-(2-Amino-1-hydroxyethyl)-5-hydroxy-2H-benzo[b][1,4]oxazin-3(4H)-one acetate, isomer 1

8-(2-azido-1-hydroxyethyl)-5-(benzyloxy)-2H-benzo[b][1,4]oxazin-3(4H)-one, isomer 1 (Example 11, step g) (0.67 g) was stirred vigorously with a mixture of 10% carbon-palladium catalyst (0.210 g) in ethanol (30 mL) under a hydrogen pressure of 4 bar for 18 hours. The catalyst was filtered off and the solvent was evaporated under reduced pressure. The residue was dissolved in acetic acid (15 mL) and EtOAc (15 mL). The mixture was filtered, and fresh 10% carbon-on-palladium catalyst (0.210 g) was added and stirred under a hydrogen pressure of 4 bar for 18 hours. The mixture was filtered, and fresh 10% carbon-on-palladium catalyst (0.210 g) was added and stirred under a hydrogen pressure of 4 bar for 18 hours. The mixture was filtered and the solvent was removed under reduced pressure. The residue was triturated with acetonitrile (30 mL). The yield was 0.33 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 6.85 (d, 1H), 6.50 (d, 1H), 4.74 - 4.69 (m, 1H), 4.52-4.42 (m, 2H), 2.74 - 2.50 (m) , 2H), 1.83 (s, 3H). Six unobserved exchangeable protons.

i) 5-hydroxy-8-(1-hydroxy-2-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Undecane-9-yl)methyl)phenethylamino)ethyl)-2H-benzene well [b][1,4]oxazin-3(4H)-one di-trifluoroacetate, Isomer 1

(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-methylthiazole-4 - ketone ketone (Example 6, step b) (0.198 g) in DCM (7 mL) was cooled in ice-bath with trifluoroacetic acid (0.037mL) followed by Dess-Martin periodinane (303mg )deal with. The mixture was stirred at 20 ° C for 30 minutes. Additional Dess-Martin periodinane (303 mg) was added and stirred for an additional 30 minutes. The mixture was treated with a saturated aqueous solution of sodium thiosulfate (7 mL) and a saturated sodium hydrogen carbonate solution (7 mL). The mixture was extracted with EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (2 mL) and added dropwise to 8-(2-amino-1-hydroxyethyl)-5-hydroxy-2H-benzo[b][1,4]oxazine-3 (4H - ketoacetate, isomer 1 (Example 11, step h) (110 mg) in methanol (5 mL) cooled to 0 ° C in EtOAc (EtOAc) Medium 1 M, 0.387 mL), and then treated with sodium cyanoborohydride (37 mg). The reaction mixture was stirred at 20 ° C for 2 hours. The mixture was evaporated to a volume of 3 mL and partitioned between ethyl acetate (30 mL) and aqueous phosphate buffer (pH = 7.2) (50 mL). The aqueous layer was acidified by the addition of acetic acid and passed through a 10 g SCX column. The column was washed with water and subsequently flushed with 7N ammonia in methanol to dissolve the product. The solvent was evaporated under reduced pressure. The crude product was dissolved in MeOH (EtOAc) (EtOAc) The residue was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 5-35% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.059 g.

m/z622(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 9.46 (s, 1H), 7.90 (s, 1H), 7.46-7.30 (m, 4H), 6.93-6.89 (m, 1H), 6.58- 6.55 (m, 1H), 5.13-5.05 (m, 1H), 4.52 (s, 2H), 4.28 (s, 2H), 3.70 (s, 4H), 3.63 (s, 2H), 3.29-2.98 (m, 10H), 2.68 (s, 3H), 2.09-1.95 (m, 2H), 1.85-1.70 (m, 2H). Five unobserved exchangeable protons.

Example 12 (R)-5-(2-(3-((4-(5-ethylthio)-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)methyl)phenethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetate

a) 3-tertiaryoxy-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester

Chloroacetate chloride (4.88 mL) was added dropwise to potassium carbonate (17.43 g) in water (78 mL) and 4-(aminomethyl)-4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (10.4 g) The mixture was vigorously stirred at 0 ° C in ethyl acetate (92 mL). After 30 minutes at 0&lt;0&gt;C, the mixture was extracted with ethyl acetate. The residue was dissolved in THF (EtOAc) (EtOAc)EtOAc. The mixture was cooled to room temperature and allowed to stir for 18 hours. Most of the solvent was removed under reduced pressure and the residue was partitioned between ethyl acetate and brine. The residue was purified by trituration with diethyl ether (30 mL)EtOAcEtOAc The yield was 8.20 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 7.95 (s, 1H), 3.97 (s, 2H), 3.72-3.62 (m, 2H), 3.10 (d, 2H), 3.05 - 2.93 (m, 2H) ), 1.77-1.69 (m, 2H), 1.53-1.43 (m, 2H), 1.40 (s, 9H).

b) 1-oxo-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester

3-tertiaryoxy-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester (Example 12, step a) (8.2 g) in THF (100 mL The solution was treated with a borane THF complex (1M in THF, 91 mL). Borane dimethyl sulfide complex (2M in THF, 15.17 mL) was added and the resulting mixture was heated at 55 °C for 2 h. The mixture was cooled to room temperature and quenched with methanol, then the solvent was evaporated under reduced pressure. The residue was dissolved in methanol (250 mL), to a solution of N1, N2 - dimethyl-ethane-1,2-diamine (10g) processing, and forming the mixture was heated at reflux for 6 hours. Additional N1,N2 -dimethylethane-1,2-diamine (3 g) was added and heating was continued under reflux for 6 hours. The mixture was cooled to room temperature and the solvent was evaporated evaporated. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 7.40 g.

_{^{1 HNMR (400MHz, CDCl 3)}} δ3.72 (s, 2H), 3.68-3.64 (m, 2H), 3.14 (t, J = 20Hz, 2H), 2.87-2.81 (m, 2H), 2.68 (s, 2H), 1.97-1.88 (m, 2H), 1.46 (s, 9H), 1.44-1.36 (m, 2H). An unobserved exchangeable proton.

c) 4-(2,2,2-trifluoroethenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester

A solution of trifluoroacetic anhydride (0.496 mL) in DCM (3 mL) was added dropwise over a 5 min period to triethylamine (0.538 mL) and 1-oxa-4,9-diazaspiro[ 5.5] Decantane-9-carboxylic acid tert-butyl ester (Example 12, step b) (0.9 g) in DCM (20 mL) EtOAc. The resulting solution was stirred at 0 ° C for 30 minutes. Water (20 mL) was added and the mixture was stirred vigorously for 10 min. The organic layer was separated, dried over sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on silica gel eluting with 40% ethyl acetate in isohexane. The pure fractions were evaporated to dryness to give the subtitle compound. The yield is 1g. Used directly without purification.

d) 2,2,2-trifluoro-1-(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)ethanone trifluoroacetate

Trifluoroacetic acid (15 mL) was added to 4-(2,2,2-trifluoroethenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid Butyl ester (Example 12, step c) (1 g) in a stirred solution of DCM (15 mL) at 20 °. The resulting solution was stirred at 20 ° C for 10 minutes. Toluene (50 mL) was added and the solvent was evaporated <RTI ID=0.0> The yield was 1.4g.

m/z 253(M+H) ⁺ (APCI)

e) 2,2,2-trifluoro-1-(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane -4-yl) ethyl ketone

A solution of 2-(3-(bromomethyl)phenyl)ethanol (Example 6, step a) (0.223 g) in acetonitrile (1 mL) was added dropwise to triethylamine (0.318 mL) over 2 min. And 2,2,2-trifluoro-1-(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)ethanone trifluoroacetate (Example 12, steps d) (0.38 g) in a stirred solution of acetonitrile (2 mL) at 20 °C. The resulting mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated and the residue was crystalljjjjjjjjjjjjjjjjj The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.260 g.

m/z 387(M+H) ⁺ (APCI)

f) (R)-5-(1-(Third-butyldimethylmethylalkyloxy)-2-(3-((4-(2,2,2-trifluoroethenyl)-1) -oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)-8-hydroxyquinoline-2(1H)-one

Adding trifluoroacetic acid (0.052 mL) to 2,2,2-trifluoro-1-(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diaza Spirulina [5.5] undecane-4-yl)ethanone (Example 12, step e) (0.26 g) in dichloromethane (7.0 mL) EtOAc. The mixture was treated with Dess-Martin periodinane (0.428 g) and stirred at 20 ° C for 30 minutes. The mixture was treated with a saturated aqueous solution of sodium thiosulfate (7.0 mL) and saturated sodium hydrogen carbonate (s. The mixture was extracted with EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (7.0mL), the (R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8-hydroxy-quinolin-2 (1H)-ketone (WO 2004106333) (0.225 g) and acetic acid (0.039 mL) were added to a mixture cooled at 0 °C. Sodium triethoxysulfonium borohydride (0.214 g) was added and the resulting mixture was stirred at 20 ° C for 90 minutes. Most of the methanol was evaporated, and the residue was partitioned between ethyl acetate and aqueous phosphates (pH = 7.2). The organic layer was dried over sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on silica gel eluting with 1% aqueous concentrated aqueous ammonia and 10% methanol in dichloromethane. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.144 g.

m/z 703(M+H) ⁺ (APCI)

g) (R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl (3-((4-(2,2, 2-Trifluoroethenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethyl)carbamic acid tert-butyl ester

Triethylamine trihydrofluoride (0.044 mL) was dissolved in methanol (1 mL) was added to a solution of the (R) -5- (1- (third - butyldimethylsilyl silicon alkyloxy) -2- ( 3-((4-(2,2,2-Trifluoroethenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenyl Amino)ethyl)-8-hydroxyquinoline-2(1H)-one (Example 12, step f) (0.140 g) in THF (4 mL). The resulting solution was stirred at 20 ° C for 18 hours. The reaction mixture was treated with triethylamine (0.111 mL) then EtOAc (EtOAc) The mixture was stirred at 20 ° C for 3 hours and then the solvent was evaporated under reduced pressure. The residue was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.100 g.

m/z 689(M+H) ⁺ (APCI)

h) (R)-3-(1-oxa-4,9-diazaspiro[5.5]undecane-9-ylmethyl)phenethyl (2-hydroxy-2-(8-hydroxyl) -2-Sideoxy-1,2-dihydroquinolin-5-yl)ethyl)carbamic acid tert-butyl ester

(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl (3-((4-(2,2,2-) Trifluorobutyl trifluoroethenyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl)phenethyl)carbamic acid (Example 12) A solution of the step g) (0.100 g) in EtOAc (EtOAc) (EtOAc) Methanol was distilled off under a stream of nitrogen and additional water (5.0 mL) was added. The solution was acidified by the addition of acetic acid and passed through 10 g of C18 cerium oxide column. The column was washed with water and then rinsed with methanol to remove the product. The solvent was distilled off under reduced pressure to give EtOAc. This solid was dissolved in methanol (2mL) and 0.700g of pass through VARIAN Bond Elut NH ₂ resin and the solvent was distilled off under reduced pressure, to produce the sub-title compound. The yield was 0.074 g.

m/z 593(M+H) ⁺ (APCI)

i) (R)-5-(2-(3-((4-(5-ethylthiophen-2-yl))-1-oxa-4,9-diazaspiro[5.5]undecane -9-yl)methyl)phenethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetate

(R)-3-(1-oxa-4,9-diazaspiro[5.5]undecane-9-ylmethyl)phenethyl (2-hydroxy-2-(8-hydroxy-2) a solution of 3-butoxy-1,2-dihydroquinolin-5-yl)ethyl)carbamic acid tert-butyl ester (Example 12, step h) (0.068 g) in DMF (2.0 mL) Triethylamine (0.048 mL) followed by 5-ethylthiophene-2-carboxylic acid (0.018 g) eluted with HATU (0.057 g). The mixture was stirred at 20 ° C for 2 hours. A "880" aqueous ammonia solution (4 drops) was added and stirred for 1 hour. The mixture was partitioned between EtOAc and EtOAc. The residue was dissolved in dichloromethane (1 mL). The solution was allowed to stand at 20 ° C for 10 minutes, toluene (20 mL) was added, and the solvent was evaporated under reduced pressure. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.039 g.

m/z 631 (M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D _{6 -} DMSO, 90 ° C) δ 8.18 (d, J = 10.0 Hz, 1H), 7.46 - 7.33 (m, 4H), 7.23(d,1H), 7.14(d,1H), 7.00(d,1H), 6.84(d,1H),6.55(d,1H),5.39-5.33(m,1H), 4.29(s,2H) , 3.74-3.63 (m, 4H), 3.54 (s, 2H), 3.29 (t, 2H), 3.21-3.00 (m, 8H), 2.83 (q, 2H), 2.10 - 1.96 (m, 2H), 1.84 -1.69 (m, 2H), 1.26 (td, 3H). Six unobserved exchangeable protons.

Example 13 5-hydroxy-8-(1-hydroxy-2-(3-((4-(2-methylthiazol-4-yl))-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)ethyl)-2H-benzo[b][1,4]oxazine-3(4H)-one di-trifluoroacetate, different Structure 2

a) 8-(2-azido-1-indenylethyl)-5-(benzyloxy)-2H-benzo[b][1,4]oxazin-3(4H)-one, Isomer 2

Isomer 2 is the second isolating isomer obtained from palmitic HPLC separation as detailed in step g of Example 11. The fractions containing the second isomerized isomer are combined and concentrated in vacuo to give the sub-title compound. The yield was 0.72 g.

m/z 339(MH) ^- (APCI)

b) (S)-8-(2-Amino-1-hydroxyethyl)-5-hydroxy-2H-benzo[b][1,4]oxazin-3(4H)-one acetate, different Structure 2

8-(2-azido-1-hydroxyethyl)-5-(benzyloxy)-2H-benzo[b][1,4]oxazin-3(4H)-one, isomer 2 (Example 13, step a) (0.72 g) A solution of a mixture of acetic acid (10 mL) and ethanol (10 mL) was vigorously stirred with a 10% carbon-on-palladium catalyst (0.225 g) under a hydrogen pressure of 4 bar for 18 hours. . The mixture was filtered, and fresh 10% carbon-on-palladium catalyst (0.225 g) was added and stirred under a hydrogen pressure of 4 bar for 18 hours. The mixture was filtered, and fresh 10% carbon-on-palladium catalyst (0.225 g) was added and stirred under a hydrogen pressure of 4 bar for 18 hours. The mixture was filtered and the solvent was removed under reduced pressure. The residue was triturated with acetonitrile (30 mL). The yield was 0.28 g.

m/z 222(MH) ^- (APCI)

c) 5-hydroxy-8-(1-hydroxy-2-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Undecane-9-yl)methyl)phenethylamino)ethyl)-2H-benzene well [b][1,4]oxazin-3(4H)-one di-trifluoroacetate, Isomer 2

(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-methylthiazole-4 a solution of the ketone (meth) ketone (Example 6, step b) (0.163 g) in EtOAc (EtOAc) (EtOAc) 0.246 g) treatment. The mixture was stirred at 20 ° C for 30 minutes, treated with additional Dess-Martin periodinane (0.246 g) and stirred at 20 ° C for an additional 30 minutes. The mixture was treated with a saturated aqueous solution of sodium thiosulfate (7.0 mL) and saturated sodium bicarbonate (7.0 mL) and stirred vigorously for 10 min. The mixture was extracted with EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (2.0 mL) and added dropwise to 8-(2-amino-1-hydroxyethyl)-5-hydroxy-2H-benzo[b][1,4]oxazine-3 ( 4H)-ketoacetate, isomer 2 (Example 13, step b) (0.090 g) in methanol (5.0 mL) cooled to 0 ° C, and with acetic acid (0.018 mL) then HCl (1M in diethyl ether, 0.32 mL), and then taken with sodium cyanoborohydride (0.030 g). The mixture was stirred at 20 ° C for 2 hours. The mixture was evaporated to a volume of 3 mL and partitioned between ethyl acetate (30 mL) and aqueous phosphate buffer (pH = 7.2) (50 mL). The aqueous layer was acidified by the addition of acetic acid and passed through a 10 g SCX column. The column was washed with water and subsequently flushed with 7N ammonia in methanol to dissolve the product. The solvent was evaporated under reduced pressure. The crude product was dissolved in MeOH (EtOAc) (EtOAc) The residue was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 5-35% acetonitrile). The fractions containing the desired product were evaporated to dryness to give the title compound. The yield was 0.053 g.

m/z 622 (M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D _{6 -} DMSO, 90 ° C) δ 9.45 (s, 1H), 7.90 (d, J = 4.6 Hz, 1H), 7.45- 7.31 (m, 4H), 6.95-6.88 (m, 1H), 6.60-6.54 (m, 1H), 5.14-5.07 (m, 1H), 4.54 (s, 2H), 4.30 (s, 2H), 3.70 ( s, 4H), 3.64 (s, 2H), 3.28-2.98 (m, 10H), 2.68 (s, 3H), 2.08-1.97 (m, 2H), 1.88-1.71 (m, 2H). Five unobserved exchangeable protons.

Example 14 (R)-8-hydroxy-5-(1-hydroxy-2-(2-(5-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diaza) Hetero[5.5]undecane-9-yl)methyl)thiophen-2-yl)ethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

a) 5-(2-(Third-butyldimethylmethylalkyloxy)ethyl)thiophene-2-carbaldehyde

Tri -butyldimethylbenzyl chloride (2.82 g) was added portionwise at 20 ° C over 20 min to 2-(thiophen-2-yl)ethanol (2 g) and imidazole (1.275 g) in DMF (20 mL) In the stirred solution. The mixture was stirred at 20 ° C for 18 hours and then EtOAc (EtOAc)EtOAc. The residue was purified by flash chromatography on EtOAc to elute from 1 to 4% ethyl acetate in isohexane. The residue (2.7 g) was dissolved in THF (50 mL), and the solution was cooled to -78 ° C, n-butyllithium (2.5 M, 5.06 mL in hexane) was added dropwise over 10 min and the mixture was stirred at 0 ° C 1 hour. The reaction mixture was cooled to -78 ° C and DMF (5.7 mL) was added dropwise over 5 min. The mixture was stirred at 20 ° C for 3 hours. The reaction mixture was partitioned between water and ethyl acetate. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 2.3 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ9.83 (s, 1H), 7.61 (d, 1H), 6.96 (d, 1H), 3.86 (t, 2H), 3.07 (t, 2H), 0.88 (s, 9H), 0.02 (s, 6H).

b) (9-((5-(2-(Terve-butyldimethylsilyloxy)ethyl)thiophen-2-yl)methyl)-1-oxa-4,9-diaza Hetero[5.5]undecane-4-yl)(2-methylthiazol-4-yl)methanone

(2-methylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 4, step h (0.25 g) and 5-(2-( Third -butyldimethylsilyloxy)ethyl)thiophene-2-carbaldehyde (Example 14, step a) (0.171 g) in NMP (5 mL) The solution in acetic acid (0.036 mL) was treated with sodium <RTI ID=0.0> The mixture was partitioned between EtOAc EtOAc m. The yield was 0.3 g.

m/z 536(M+H) ⁺ (APCI)

c) (9-((5-(2-hydroxyethyl)thiophen-2-yl)methyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl (2-methylthiazol-4-yl)methanone

(9-((5-(2-( Third -butyldimethylsilyloxy)ethyl)thiophen-2-yl)methyl)-1-oxa-4,9-diazaspiro [5.5] undecane-4-yl)(2-methylthiazol-4-yl)methanone (Example 14, step b) (0.3 g) in THF (5 mL) Treatment with tetrabutylammonium solution (1M in THF, 0.672 mL). The mixture was allowed to stand at 20 ° C for 30 minutes. The solvent was evaporated using a stream of nitrogen and the residue was purified by flash chromatography eluting with EtOAc EtOAc. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.187 g.

m/z 422(M+H) ⁺ (APCI)

d) (R)-5-(1-(Third-butyldimethylmethylalkyloxy)-2-(2-(5-((4-(2-methylthiazole-4-carbonyl))-) 1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)thiophen-2-yl)ethylamino)ethyl)-8-hydroxyquinoline-2 (1H)-ketone

(9-((5-(2-hydroxyethyl)thiophen-2-yl)methyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)( 2-methylthiazol-4-yl)methanone (Example 14, step c) (0.18 g) in dichloromethane (10 mL) at 20 ° C with trifluoroacetic acid (0.033 mL) Martin was treated with iodosane (0.254 g) and the mixture was stirred at 20 ° C for 90 minutes. The mixture was then treated with saturated aqueous sodium thiosulfate (10 mL) and saturated sodium bicarbonate (10 mL) and stirred vigorously for 10 min. The mixture was extracted with EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (2mL) and added to (R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8-hydroxy-quinolin-2 (1H)-ketone (WO 2004106333) (0.143 g) and acetic acid (0.024 mL) in methanol (7 mL). The mixture was cooled in an ice-bath and sodium triethyl succinyl borohydride (0.136 g). The ice bath was removed and the mixture was stirred at room temperature for 1 hour. Most of the methanol was evaporated and the residue was crystalljjjjjjjjjjjjjj The crude product was purified by flash chromatography on silica gel eluting with 9% methanol and 1% ' The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.078 g.

m/z 738(M+H) ⁺ (APCI)

e) (R)-8-hydroxy-5-(1-hydroxy-2-(2-(5-((4-(2-methylthiazol-4-yl))-1-oxa-4,9 -diazaspiro[5.5]undecane-9-yl)methyl)thiophen-2-yl)ethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

( R )-5-(1-( Third -butyldimethylmethylalkyloxy)-2-(2-(5-((4-(2-methylthiazole-4-carbonyl)-1-) Oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)thiophen-2-yl)ethylamino)ethyl)-8-hydroxyquinoline-2 (1H a solution of the ketone (Example 14, step d) (0.078 g) in a mixture of THF (4 mL) and methanol (1 mL) with triethylamine trihydrofluorate (0.022 mL) and mixture Leave at °C for 18 hours. The solvent was evaporated under reduced pressure and the residue was azeotroped with toluene. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 10 to 30% acetonitrile). The fractions containing the desired product were evaporated to dryness to give the title compound. The yield was 0.021 g.

m / z 624 (M + H ) + (APCI) 1 H NMR (400MHz, D 6 -DMSO) δ10.50 (s, 2H), 10.15-9.84 (m, 1H), 8.91 (s, 1H), 8.80 (s, 1H), 8.16 (d, J = 10.0 Hz, 1H), 8.00 (s, 1H), 7.26-7.12 (m, 2H), 7.02-6.94 (m, 2H), 6.58 (d, J = 9.7 Hz,1H), 6.21(s,1H), 5.36-5.31(m,1H),4.53(s,2H),3.83-3.48(m,6H),3.33-2.93(m,10H),2.68(s, 3H), 2.16-2.04 (m, 2H), 1.79-1.59 (m, 2H).

Example 15 (R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one trifluoroacetate

Hydrogen chloride (2M in diethyl ether, 0.504 mL) was added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one acetate (Example 1, step d) (0.222 g) and 2-(3-((4-(5-methylthiophen-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5] A solution of the undecane-9-yl)methyl)phenyl)acetaldehyde (Example 9, step d) (0.320 g) in methanol (10 mL). After 5 min, sodium cyanoborohydride (0.100 g) was added and the mixture was stirred overnight. The solution was concentrated to 3 mL and partitioned between pH 7.2 buffer (80 mL) and ethyl acetate. A saturated sodium bicarbonate solution was added to the aqueous layer which was then extracted with ethyl acetate. The combined ethyl acetate solution was dried over sodium sulfate, filtered and evaporated to dryness. Purification by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in acetonitrile 25-65%). The fractions containing the product were combined and evaporated to dryness in vacuo. Acetonitrile was added and the solution was evaporated in vacuo and the procedure was repeated. Diethyl ether was added and the gum was triturated to give the title compound as a solid. The yield was 0.017 g.

m / z 623 (M + H ) + (APCI) 1 H NMR (400MHz, D 6 -DMSO) δ11.67 (s, 1H), 10.25 (s, 1H), 9.73-9.63 (m, 1H), 8.99 -8.89 (m, 1H), 8.85-8.76 (m, 1H), 7.46-7.32 (m, 3H), 7.30-7.20 (m, 1H), 6.93 (d, J = 8.3 Hz, 1H), 6.86-6.81 (m, 1H), 6.77 (d, J = 8.3 Hz, 1H), 6.53-6.45 (m, 1H), 4.93-4.86 (m, 1H), 4.45-4.39 (m, 0.4H), 4.35-4.28 ( m, 1.6H), 3.74-3.60 (m, 4H), 3.49 (s, 2H), 3.27-3.14 (m, 4H), 3.13-2.89 (m, 6H), 2.46 (s, 3H), 2.12-2.03 (m, 2H), 1.70-1.59 (m, 2H). An unobserved exchangeable proton.

Example 16 (R)-4-hydroxy-7-(1-hydroxy-2-(4-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(4-(bromomethyl)phenyl)ethanol

Borane dimethyl sulfide complex (2M in THF, 4.37 mL) was added dropwise to 2-(4-(bromomethyl)phenyl)acetic acid (1.0 g) in THF (20 mL) The solution was stirred and the mixture was stirred for 1 hour. Methanol was added dropwise until bubbling ceased and the solvent was evaporated in vacuo. Purified by silica gel chromatography to dissolve in 30% ethyl acetate in isohexane. The fractions containing the product were combined and evaporated in vacuo to give a white crystalline solid. The yield was 0.86 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ7.36-7.32 (m, 2H), 7.20 (d, J = 8.2Hz, 2H), 4.68 (s, 2H), 4.64 (t, J = 5.1Hz, 1H), 3.64-3.55 (m, 2H), 2.75-2.67 (m, 2H).

b) (9-(4-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-methylthiazole -4-yl)methanone

(2-methylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 4, step h (0.791g) and a solution of triethylamine (0.697mL) in acetonitrile (10mL) were added 2-(4-(bromomethyl)phenyl)ethanol (Example 16, step a) and the resulting mixture was stirred. Overnight. The solvent was evaporated in vacuo. The residue was partitioned between EtOAc (25 mL)EtOAcEtOAc. The aqueous phase was separated and extracted with ethyl acetate (2×25 mL). The combined organics were washed with EtOAc (EtOAc)EtOAc. The yield was 0.76 g.

m/z 416(M+H) ⁺ (APCI)

c) 2-(4-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl Phenyl) acetaldehyde

Add TFA (0.030 mL) to (9-(4-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl) (2-Methylthiazol-4-yl)methanone (Example 16, step b) (0.16 g) in DCM (5 mL) in EtOAc. Dess-Martin periodinane (0.245 g) was added and the mixture was allowed to warm to room temperature and stirred for 45 min. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine. AcOH (0.1 mL) was then added, and the mixture was dried over sodium sulfate, filtered and evaporated in vacuo to give the title compound. The yield was 0.16 g.

d) (R)-4-hydroxy-7-(1-hydroxy-2-(4-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

2-(4-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl)benzene A solution of acetaldehyde (Example 16, step c) (0.08 g) in methanol (2 mL) was added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzene. [d] A mixture of thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.076 g) and acetic acid (0.017 mL) in methanol (0.5 mL). The resulting mixture was stirred for 5 min and then cooled to 0 °C. Sodium cyanoborohydride (0.012 g) was then added, and the mixture was allowed to warm to room temperature and stirred for 2 hr. The reaction mixture was applied to a C18 column (Varian 10 g). The column was washed with water (50 mL) and EtOAc (50 mL). Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile). The title compound was obtained as a white solid. The yield was 0.66 g.

m/z 624 (M+H) ⁺ (APCI) ¹ H NMR (300 MHz, D _{6 -} DMSO, 90 ° C) δ 11.28 (s, 1H), 7.90 (s, 1H), 7.48 (d, J = 7.1 Hz, 2H), 7.34 (d, J = 7.3 Hz, 2H), 6.94 (d, J = 8.1 Hz, 1H), 6.78 (d, J = 8.1 Hz, 1H), 4.99 - 4.88 (m, 1H), 4.31 (s, 2H), 3.76-3.55 (m, 6H), 3.32-2.91 (m, 10H), 2.67 (s, 3H), 2.10- 1.67 (m, 4H). Five unobserved exchangeable protons.

Example 17 (R)-8-hydroxy-5-(1-hydroxy-2-(4-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

2-(4-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl)benzene A solution of acetaldehyde (Example 16, step c) (0.08 g) in methanol (2 mL) was added to ( R )-5-(2-amino-1-(tri-butyl-dimethyl decane) A mixture of methoxy)ethyl)-8-hydroxyquinolin-2(1H)-one (WO 2004106333) (0.097 g) and acetic acid (0.017 mL) in methanol (0.5 mL). The resulting mixture was stirred for 5 min and then cooled to 0 °C. Sodium cyanoborohydride (0.018 g) was then added, and the mixture was allowed to warm to room temperature and stirred for 2 hr. The reaction mixture was applied to a C18 column (Varian 10 g). The column was washed with water (50 mL) and EtOAc (50 mL). The fractions were combined, evaporated and purified by gel chromatography, eluting with 95:5:0.5 to 89:10:1 DCM:methanol: '880' ammonia solution. The residue was dissolved in THF (2 mL) EtOAc (EtOAc) Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile). The title compound was obtained as a white solid. The yield was 0.07 g.

m/z 618 (M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D _{6 -} DMSO, 90 ° C) δ 8.18 (d, J = 9.7 Hz, 1H), 7.90 (s, 1H), 7.48 ( d, J = 7.7 Hz, 2H), 7.34 (d, J = 7.9 Hz, 2H), 7.14 (d, J = 8.2 Hz, 1H), 7.00 (d, J = 7.9 Hz, 1H), 6.54 (d, J = 10.0 Hz, 1H), 5.36 (dd, J = 8.6, 4.0 Hz, 1H), 4.31 (s, 2H), 3.75-3.58 (m, 6H), 3.33 - 2.97 (m, 10H), 2.67 (s , 3H), 2.11-1.67 (m, 4H). Six unobserved exchangeable protons.

Example 18 (R)-7-(2-(2,5-Dimethyl-4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)ethoxy)benzylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoro Acetate

a) 4-hydroxy-2,5-dimethylbenzoic acid

A solution of boron tribromide (1M in DCM, 7.21 mL) was added dropwise to a suspension of 4-methoxy-2,5-dimethylbenzoic acid (1 g) in DCM (5 mL) at -78. The reaction was allowed to warm to RT and stirred overnight. The reaction was cooled to -78 ° C and a solution of boron tribromide (1M in DCM, 7.. The reaction was allowed to warm to RT and stirred overnight. The reaction was carefully poured onto ice (~50 mL). The resulting aqueous solution was extracted with DCM (5×50 mL). The combined organic solution was washed with EtOAc EtOAc m. The yield was 0.75 g.

¹ H NMR (300 MHz, D ₆ -DMSO) δ 12.19 (s, 1H), 9.91 (s, 1H), 7.64 (s, 1H), 6.65 (s, 1H), 2.43 (s, 3H), 2.09 ( s, 3H).

b) Methyl 4-hydroxy-2,5-dimethylbenzoate

Acetyl chloride (0.107 mL) was added dropwise to a solution of 4-hydroxy-2,5-dimethylbenzoic acid (Example 18, step a) (0.249 g) in methanol (15 mL) Stir at room temperature for 72 h. Ethyl chloride (0.107 mL) was then added and the reaction was heated to 50 ° C and stirred at this temperature overnight. The reaction was concentrated and the residue was purified eluting eluting eluting eluting The fractions containing the product were combined and evaporated in vacuo to give a sub-title compound as a white solid. The yield is 0.25 g.

_{^{1 H NMR (300MHz, D 6}} -DMSO) δ10.04 (s, 1H), 7.64 (s, 1H), 6.67 (s, 1H), 3.74 (s, 3H), 2.42 (s, 3H), 2.09 ( s, 3H).

c) Methyl 4-(2,2-diethoxyethoxy)-2,5-dimethylbenzoate

Add 2-bromo-1,1-diethoxyethane (0.31 mL) to methyl 4-hydroxy-2,5-dimethylbenzoate (Example 18, step b) (0.25 g) and carbonic acid铯 (0.68 g) in a mixture of DMF (20 mL). The resulting mixture was stirred at 85 ° C for 16 h. The reaction was partitioned between water (200 mL) and ethyl acetate (100 mL). The aqueous phase was separated and extracted with ethyl acetate (2×100 mL). The combined organic solution was washed with water (2×100 mL) and brine (100 mL). The residue was purified by EtOAc (EtOAc) elut elut The fractions containing the product are combined and evaporated in vacuo to give the sub-title compound as a clear oil. The yield was 0.26 g.

_{^{1 H NMR (300MHz, CDCl 3}} ) δ7.75 (s, 1H), 6.64 (s, 1H), 4.86 (t, J = 5.2Hz, 1H), 4.04 (d, J = 5.4Hz, 2H), 3.88 -3.58 (m, 7H), 2.58 (s, 3H), 2.20 (s, 3H), 1.25 (t, J = 7.0 Hz, 6H).

d) (4-(2,2-diethoxyethoxy)-2,5-dimethylphenyl)methanol

A lithium aluminum hydride solution (1 M in THF, 1.17 mL) was added dropwise to methyl 4-(2,2-diethoxyethoxy)-2,5-dimethylbenzoate (Example 18, step c) (0.231 g) in THF (20 mL) in EtOAc. The resulting mixture was allowed to warm to RT and stirred for 1 h. Ethanol (0.5 mL) was added cautiously and the reaction was concentrated. The residue was partitioned between 1M EtOAc (50 mL) andEtOAc. The aqueous solution was separated and extracted with ethyl acetate (2×50 mL). The combined organics were washed with EtOAc (EtOAc)EtOAc. The yield was 0.20 g.

_{^{1 H NMR (300MHz, CDCl 3}} ) δ7.08 (s, 1H), 6.66 (s, 1H), 4.85 (t, J = 5.3Hz, 1H), 4.60 (s, 2H), 4.00 (d, J = 5.2 Hz, 2H), 3.84-3.58 (m, 4H), 2.33 (s, 3H), 2.20 (s, 3H), 1.25 (t, J = 7.0 Hz, 6H). An unobserved exchangeable proton.

e) (9-(2-(4-(hydroxymethyl)-2,5-dimethylphenoxy)ethyl)-1-oxa-4,9-diazaspiro[5.5] eleven Cycloalkyl-4-yl)(2-methylthiazol-4-yl)methanone

(4-(2,2-diethoxyethoxy)-2,5-dimethylphenyl)methanol (Example 18, step d) (0.2 g) in acetic acid (5 mL) and water (5 mL) The solution in the mixture was heated at 65 °C for 1 h and allowed to cool to RT. The reaction mixture was poured into EtOAc EtOAc (EtOAc) The combined organic solution was washed with brine (25 mL) dried over sodium sulfate. The residue was redissolved in methanol (5 mL) and acetic acid (5 mL) was added followed by (2-methylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane- 4-yl)methanone trifluoroacetate (Example 4, step h) (0.35 g). The resulting mixture was stirred for 15 min then sodium cyanoborohydride (0.07 g) was added and the mixture was stirred overnight. The solvent was evaporated and the residue was crystalljjjjjjjjjjj The layers were separated and the aqueous extracted with ethyl acetate (2×30 mL). The combined organic solution was washed with brine (20 mL) dried over sodium sulfate. The residue was purified by EtOAc EtOAc (EtOAc:EtOAc:EtOAc: The fractions containing the product were combined and evaporated in vacuo to give a sub-title compound. The yield was 0.12 g.

m/z 460(M+H) ⁺ (APCI)

f) 2,5-Dimethyl-4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane -9-yl)ethoxy)benzaldehyde

Manganese dioxide (0.32 g) was added to (9-(2-(4-(hydroxymethyl)-2,5-dimethylphenoxy)ethyl)-1-oxa-4,9-di A solution of azaspiro[5.5]undec-4-yl)(2-methylthiazol-4-yl)methanone (Example 18, step e) (0.17 g) in DCM (10 mL). The resulting mixture was heated under reflux for 4 h. The reaction was filtered through celite and the pad was washed with DCM (2.times.50mL). The mother liquor and the wash solution are combined and evaporated to yield a yellow gum of the sub-title compound. The yield was 0.17 g.

m/z 458(M+H) ⁺ (APCI)

g) (R)-7-(2-(2,5-Dimethyl-4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)ethoxy)benzylamino)-1-hydroxyethyl)-4-mercaptobenzo[d]thiazole-2(3H)-one -trifluoroacetate

2,5-Dimethyl-4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 9-yl)ethoxy)benzaldehyde (Example 18, step f) (0.08 g) was added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d a mixture of thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.07 g) and acetic acid (0.010 mL) in methanol (2 mL). The mixture was stirred for 30 min and then cooled in an ice bath. Sodium cyanoborohydride (0.016 g) was then added and the mixture was stirred for 2 h. The solvent was evaporated in vacuo and EtOAcqqqqqqqqqq The aqueous solution was separated and extracted with ethyl acetate (2×50 mL). The combined organic solution was washed with brine (20 mL) dried over sodium sulfate. The residue was purified by silica gel chromatography eluting with a gradient of 95:5:0.5 to 92:8:0.8 DCM: methanol: ' The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile). The title compound was obtained as a white solid. The yield was 0.069 g.

m/z 668(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.34-11.14 (m, 1H), 7.92 (s, 1H), 7.26 (s, 1H), 6.93 (d, J = 8.2 Hz, 1H) , 6.86 (s, 1H), 6.76 (d, J = 8.2 Hz, 1H), 4.94 (t, J = 6.5 Hz, 1H), 4.39 - 4.31 (m, 2H), 4.23-4.09 (m, 2H), 3.78-3.63 (m, 6H), 3.61-3.55 (m, 2H), 3.49-3.20 (m, 4H), 3.13-3.03 (m, 2H), 2.68 (s, 3H), 2.33 (s, 3H), 2.21-2.01 (m, 5H), 1.94-1.78 (m, 2H). Five unobserved exchangeable protons.

Example 19 (R)-7-(2-(3-fluoro-5-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] eleven Cycloalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(3-(bromomethyl)-5-fluorophenyl)acetic acid

Benzoyl peroxide (0.05 g) was added to 2-(3-fluoro-5-methylphenyl)acetic acid (0.518 g) and N -bromosuccinimide (0.6 g) in DCM (10 mL) In the mixture. The reaction was heated at reflux for 1 h. DCM (10 mL) and water (20 mL) were added and the organic phase was separated. The organic layer was washed with brine (20 mL) dried over sodium sulfate. The residue was triturated with toluene and filtered to remove a white solid. The mother liquor was evaporated in vacuo to give a sub-title compound as a white solid. The yield was 0.38 g.

b) (9-(3-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -methylthiazole-4-yl)methanone

A solution of borane dimethyl sulfide complex (2M in THF, 3.85 mL) was added dropwise to 2-(3-(bromomethyl)-5-fluorophenyl)acetic acid (Example 19, step a (0.38 g) in THF (10 mL) in EtOAc. The resulting mixture was allowed to warm to RT and stirred for 1 h. The reaction was cooled to 0 ° C and methanol (1 mL) was added dropwise until bubbling ceased. The solvent was evaporated in vacuo. Subsequent addition of (2-methylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 4, Step h) (0.61 g) followed by triethylamine (0.54 mL). The solvent was then evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 99:1:0.1 to 94.5:5:0.5 DCM: methanol: '880' aqueous ammonia solution. The fractions containing the product were combined and evaporated in vacuo to give a yellow foam of sub-title compound. The yield was 0.57 g.

m/z 434(M+H) ⁺ (APCI)

c) 2-(3-Fluoro-5-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9- Methyl)phenyl)acetaldehyde

Add TFA (0.08 mL) to (9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 4-Methyl)(2-methylthiazol-4-yl)methanone (Example 19, step b) (0.46 g), m. m. Dess-Martin periodinane (0.68 g) was then added and the mixture was stirred at RT for 45 min. Saturated sodium thiosulfate solution (5 mL), saturated sodium bicarbonate (5 mL) and ethyl acetate (20 mL) The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with EtOAc (EtOAc)EtOAc. The yield is 0.4g.

m/z 432(M+H) ⁺ (APCI)

d) (R)-7-(2-(3-Fluoro-5-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

2-(3-Fluoro-5-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl) A solution of methyl (phenyl) acetaldehyde (Example 19, step c) (0.2 g) in methanol (3 mL) was added to ( R )-7-(2-amino-1-hydroxyethyl)-4 a mixture of hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.18 g) and acetic acid (0.04 mL) in methanol (0.5 mL). The resulting mixture was stirred for 5 min and then cooled to 0 °C. Sodium cyanoborohydride (0.03 g) was then added and the mixture was warmed to RT and stirred for 2 h. The solvent was evaporated in vacuo and purified by silica gel chromatography eluting with gradient: 95:5:0.5 to 89:10:1 DCM:methanol: The product-containing fractions were combined the solution was evaporated in vacuo and purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile) to borrow. The title compound was obtained as a white solid. The yield was 0.018 g.

m/z 642(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.89 (s, 1H), 7.26-7.12 (m, 3H), 6.93 (d, J = 8.5 Hz, 1H), 6.77 (d, J = 8.2 Hz, 1H), 4.94-4.85 (m, 1H), 4.26-4.13 (m, 2H), 3.74-3.59 (m, 6H), 3.35-2.94 (m, 10H), 2.67 (s, 3H), 2.06 -1.91 (m, 2H), 1.83-1.68 (m, 2H). Six unobserved exchangeable protons.

Example 20 (R)-4-hydroxy-7-(1-hydroxy-2-(3-(2-(4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)ethyl)benzylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(3-(hydroxymethyl)phenyl)ethanol

Lithium aluminum hydride (1M in diethyl ether, 5.55 mL) was added dropwise to methyl 3-(2-hydroxyethyl)benzoate (1.0 g) in tetrahydrofuran (15 mL). In solution. After 1 h, the reaction mixture was quenched with methanol and evaporated in vacuo. Purification by hydrazine chromatography, eluting with ethyl acetate: hexanes: 2:1. The yield is 0.5g.

¹ H NMR (300MHz, CDCl ₃ ) δ 7.31 (t, J = 7.7 Hz, 1H), 7.26-7.20 (m, 2H), 7.16 (d, J = 7.7 Hz, 1H), 4.67 (s, 2H) , 3.86 (t, J = 6.9 Hz, 2H), 2.87 (t, J = 6.9 Hz, 2H). Two unobserved exchangeable protons.

b) 3-(2-hydroxyethyl)benzaldehyde

Manganese (IV) (2.0 g) was added to 2-(3-(hydroxymethyl)phenyl)ethanol (Example 20, step a) (0.50 g) in DCM (10 mL). In solution. After 6 h, the reaction mixture was filtered with EtOAc EtOAc. The solution was evaporated in vacuo to give a sub-title compound. Use directly. The yield was 0.46 g.

c) 2-(3-(diethoxymethyl)phenyl)ethanol

Ammonium chloride (0.08 g) was added to 3-(2-hydroxyethyl)benzaldehyde (Example 20, step b) (0.45 g) and triethoxymethane (0.55 g) in EtOH (8 mL) In the solution, the reaction mixture was heated at 70 ° C for 3 h. After cooling, the reaction mixture was partitioned between ethyl acetate and saturated sodium bicarbonate. The ethyl acetate solution was dried over sodium sulfate, filtered and evaporated in vacuo. The yield was 0.24 g.

¹ H NMR (300MHz, CDCl ₃ ) δ 7.38-7.25 (m, 3H), 7.22 - 7.16 (m, 1H), 5.50-5.47 (m, 1H), 3.93-3.82 (m, 2H), 3.71-3.47 (m, 4H), 2.94 - 2.86 (m, 2H), 1.29 - 1.21 (m, 6H). An unobserved exchangeable proton.

d) 3-(diethoxymethyl) phenethyl methanesulfonate

Methanesulfonium chloride (0.18 g) in dichloromethane (0.2 mL) was added to 2-(3-(diethoxymethyl)phenyl)ethanol (Example 20, step c) (0.23 g) And a stirred solution of triethylamine (0.429 mL) in dichloromethane (2.5 mL). After 1 h, the reaction mixture was partitioned between ethyl acetate and saturated sodium bicarbonate. The ethyl acetate layer was washed with aq. The yield was 0.28 g.

_{^{1 H NMR (300MHz, CDCl 3}} ) δ7.40-7.29 (m, 3H), 7.19 (d, J = 7.4Hz, 1H), 5.47 (s, 1H), 4.42 (t, J = 6.9Hz, 2H) , 3.67-3.48 (m, 4H), 3.07 (t, J = 6.9 Hz, 2H), 2.86 (s, 3H), 1.24 (t, J = 6.8 Hz, 6H).

e) (9-(3-(Diethoxymethyl)phenethyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(5-A) Thiophen-2-yl)methanone

3-(Diethoxymethyl)phenethyl methanesulfonate (Example 20, step d) (0.30 g) in acetonitrile (1.5 mL) was added to (5-methylthiophen-2-yl) (1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 9, step b) (0.391 g) and triethylamine (0.415) In a stirred solution of acetonitrile (1.5 mL), the solution was heated at 70 ° C for 48 h. After cooling, the solution was evaporated in vacuo and applied to EtOAc EtOAc EtOAc (EtOAc) The yield was 0.2 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.30-7.24 (m, 3H), 7.14 (d, J = 7.6Hz, 1H), 7.11 (d, J = 3.7Hz, 1H), 6.69 (dd, J = 3.7 Hz, 1.1 Hz, 1H), 5.46 (s, 1H), 3.78-3.71 (m, 4H), 3.65-3.49 (m, 6H), 2.82-2.77 (m, 2H), 2.65-2.57 (m, 4H) ), 2.50 (s, 3H), 2.48-2.39 (m, 2H), 1.94-1.86 (m, 2H), 1.67-1.56 (m, 2H), 1.23 (t, J = 7.2 Hz, 6H).

f) 3-(2-(4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethyl) Benzaldehyde

Addic acid (2 mL) to (9-(3-(diethoxymethyl)phenethyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl (5-Methylthiophen-2-yl)methanone (Example 20, step e) (0.20 g) in EtOAc (3 mL) and water (1 mL) After 10 min, the reaction mixture was evaporated in vacuo and toluene was evaporated. The residue was dissolved in acetonitrile then evaporated in vacuo. The yield was 0.16 g.

m/z413(M+H) ⁺ (APCI)

g) (R)-4-hydroxy-7-(1-mercapto-2-(3-(2-(4-(5-methylthiophen-2-carbonyl)-1-oxa-4,9-) Diazaspiro[5.5]undecal-9-yl)ethyl)benzylamino)ethyl)benzo Id]thiazole-2(3H)-one di-trifluoroacetate

Acetic acid (0.033 mL) was added to 3-(2-(4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)ethyl)benzaldehyde (Example 20, step f) (0.17 g) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole- 2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.15 g) in a stirred solution of methanol (10 mL). After 2 min, sodium cyanoborohydride (0.10 g) was added and the mixture was stirred at room temperature overnight. The methanol solution was concentrated to ~3 mL and diluted with ethyl acetate. A gelatinous precipitate formed. Ethyl acetate was removed, washed with saturated aqueous sodium bicarbonate and brine and dried over sodium sulfate. The gum was dissolved in a small amount of methanol and added to ethyl acetate. The ethyl acetate was filtered and evaporated in vacuo to give a white solid. The solid was dissolved in methanol and purified by preparative HPLC (Sunfire ^TM, Gradient: in 15 aqueous 0.2% TFA to 50% acetonitrile) to borrow. The fractions containing the pure product were combined and evaporated to dryness. The title compound was obtained as a white solid. The yield was 0.11 g.

m/z 623(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.67 (s, 1H), 10.23 (s, 1H), 9.85-9.75 and 9.61-9.50 (m, total 1H), 9.18-8.98 (m, 2H) ), 7.45-7.31 (m, 4H), 7.29-7.24 (m, 1H), 6.89 (d, J = 7.4 Hz, 1H), 6.86-6.83 (m, 1H), 6.75 (d, J = 7.4 Hz, 1H), 6.48-6.44 (m, 1H), 4.93-4.85 (m, 1H), 4.21-4.14 (m, 2H), 3.74-3.64 (m, 4H), 3.53 (s, 2H), 3.48-3.40 ( m, 2H), 3.36-3.27 (m, 2H), 3.08-2.92 (m, 6H), 2.47 (s, 3H), 2.16-2.08 (m, 2H), 1.74-1.61 (m, 2H).

Example 21 (R)-4-hydroxy-7-(1-hydroxy-2-((5-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diaza) Heterospiro[5.5]undecane-9-yl)ethyl)thiophen-2-yl)methylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 5-(2-(Third-butyldimethylmethylalkyloxy)ethyl)thiophene-2-carbaldehyde

Tri -butyldimethylbenzyl chloride (2.82 g) was added portionwise at 20 ° C over 20 min to 2-(thiophen-2-yl)ethanol (2 g) and imidazole (1.275 g) in DMF (20 mL) In the stirred solution. The mixture was stirred at 20 ° C for 18 hours and then EtOAc (EtOAc)EtOAc. The residue was purified by flash chromatography on EtOAc to elute from 1 to 4% ethyl acetate in isohexane. The residue (2.7 g) was dissolved in THF (50 mL), and the solution was cooled to -78 ° C, n-butyllithium (2.5 M, 5.06 mL in hexane) was added dropwise over 10 min and the mixture was stirred at 0 ° C 1 hour. The reaction mixture was cooled to -78 ° C and DMF (5.7 mL) was added dropwise over 5 min. The mixture was stirred at 20 ° C for 3 hours. The reaction mixture was partitioned between water and ethyl acetate. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 2.3 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 9.83 (s, 1H), 7.61 (d, 1H), 6.96 (d, 1H), 3.86 (t, 2H), 3.07 (t, 2H), 0.88 (s) , 9H), 0.02 (s, 6H).

b) 2-(5-(diethoxymethyl)thiophen-2-yl)ethanol

5-(2-( Third -butyldimethylsilyloxy)ethyl)thiophene-2-carbaldehyde (Example 21, Step a) (1.37 g) and ammonium chloride (0.135 g) and orthoformic acid A mixture of triethyl ester (0.928 mL) in ethanol (15 mL) was evaporated. Most of the ethanol was removed under reduced pressure and the residue was partitioned between ethyl acetate and sat. sodium hydrogen carbonate. The residue was dissolved in THF (20 mL) EtOAc (EtOAc) The mixture was allowed to stand at room temperature for 30 minutes. The solvent was removed under reduced pressure and the residue was purified by flash chromatography eluting with EtOAc The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.940 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ6.85 (d, 1H), 6.73 (d, 1H), 5.64 (s, 1H), 4.79-4.74 (m, 1H), 3,62-3.43 (m , 6H), 2.87 (t, 2H), 1.13 (t, 6H).

c) 5-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethyl) Thiophene-2-carbaldehyde

A solution of 2-(5-(diethoxymethyl)thiophen-2-yl)ethanol (Example 21, step b) (0.54 g) in dichloromethane (20 mL) And the mixture was cooled in an ice bath. Then a solution of methanesulfonium chloride (0.183 mL) in dichloromethane (3 mL) was added dropwise over 2 min and the mixture was stirred at 0 ° C for one hour. The mixture was washed with water and the org. The residue was dissolved in a mixture of acetonitrile (7 mL) and DMF (1 mL), then triethylamine (0.654 mL), followed by (2-methylthiazol-4-yl)(1-oxa-4,9-diazepine Heterospiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 4, step h) (0.5 g) and heated at 70 °C for 10 hours. The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The residue was dissolved in EtOAc (5 mL)EtOAc. The mixture was allowed to stand at 0 ° C for 10 minutes. The solvent was removed under reduced pressure and the residue was azeotroped twice with toluene and twice with acetonitrile. The crude product was purified by flash chromatography on silica gel using 1% triethylamine and 2% methanol in dichloromethane. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.52 g.

m/z 420(M+H) ⁺ (APCI)

d) (R)-4-hydroxy-7-(1-hydroxy-2-((5-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9- Diazaspiro[5.5]undecane-9-yl)ethyl)thiophen-2-yl)methylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoro Acetate

( R )-7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) 0.22 g) of the solution in methanol (8 mL) was taken in acetic acid (0.041 mL), followed by 5-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxo-4,9- A solution of diazaspiro[5.5]undecane-9-yl)ethyl)thiophene-2-carbaldehyde (Example 21, step c) (0.3 g) in MeOH (2 mL). The mixture was cooled in an ice-bath and treated with sodium <RTI ID=0.0> The mixture was then stirred at room temperature for 18 hours. Most of the methanol was evaporated under reduced pressure and the residue was partitioned between THF (50 mL), brine (10 mL) and sat. The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired product were evaporated to dryness to give the title compound. The yield was 0.08 g.

m/z 630(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.27 (s, 1H), 7.92 (s, 1H), 7.14 (d, 1H), 6.94-6.88 (m, 2H), 6.76 (d, 1H), 4.91-4.85 (m, 1H), 4.37 (s, 2H), 3.70 (s, 4H), 3.66 (s, 2H), 3.40-3.30 (m, 4H), 3.26-3.07 (m, 4H) , 3.04-3.02 (m, 2H), 2.68 (s, 3H), 2.09-1.99 (m, 2H), 1.87-1.73 (m, 2H). Five unobserved exchangeable protons.

Example 22 (R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester

2-isopropylthiazole-4-carboxylic acid (1 g) and 1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester hydrochloride (WuXi PharmaTech) ( 1.71 g) The solution in DMF (30 mL) was cooled in EtOAc EtOAc (EtOAc) The ice bath was removed and the mixture was stirred at 20 ° C for 1 hour. The mixture was partitioned between EtOAc and EtOAc. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield is 2g.

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ7.93 (s, 1H), 3.71-3.63 (m, 6H), 3.51-3.44 (m, 2H), 3.35-3.26 (m, 1H), 3.18-3.10 (m, 2H), 1.74-1, 67 (m, 2H), 1.49-1.41 (m, 2H), 1.39 (s, 9H), 1.34 (d, J = 7.6HZ, 6H).

b) (2-Isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone triazide

4-(2-Isopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester (Example 22, step a (2.3 g) A solution of a mixture of dichloromethane (40 mL) and trifluoroacetic acid (10 mL) was placed at 20 ° C for 30 min. Toluene (50 mL) was added and the solvent was evaporated, then this was repeated with additional toluene (50 mL). The residue was triturated with ether. The gum was then dissolved in acetonitrile and the solvent was evaporated to give subtitle compound. The yield was 1.64 g.

m/z 310(M+H) ⁺ (APCI)

c) (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-isopropyl Thiazol-4-yl)methanone

(2-Isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (0.85 g) and a mixture of 2-(3-(bromomethyl)phenyl)ethanol (Example 6, step a) (0.432 g) in acetonitrile (10 mL). 0.616 mL) and stirred for 2 hours. The solvent was evaporated under reduced pressure and EtOAc evaporated m. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.71 g.

m/z 444(M+H) ⁺ (APCI)

d) (R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diaza) Hetero[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-isopropylthiazole- A solution of 4-yl)methanone (Example 22, step c) (0.3 g) in dichloromethane (10 mL)EtOAc (EtOAc) g), and the resulting mixture was stirred at 20 ° C for 1 hour. The reaction mixture was treated with aq. sodium sulphate (10 mL) and EtOAc (EtOAc) The mixture was extracted with EtOAc EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (2 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.2 g) and acetic acid (0.039 mL) in methanol (10 mL) in 0 ° C solution. Sodium cyanoborohydride (0.064 g) was added, and the mixture was stirred at room temperature for 18 hr. Most of the methanol was evaporated under reduced pressure and the residue was partitioned between THF (50 mL), brine (10 mL) and sat. The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired product were evaporated to dryness to give the title compound. The yield was 0.170 g.

m/z 652(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.27 (s, 1H), 7.94 (s, 1H), 7.45-7.31 (m, 4H), 6.93 (d, J = 31.1 Hz, 1H) , 6.77 (d, J = 24.6 Hz, 1H), 4.96-4.87 (m, 1H), 4.27 (s, 2H), 3.70 (s, 4H), 3.65 (s, 2H), 3.35-2.97 (m, 11H) ), 2.08-1.96 (m, 2H), 1.84-1.67 (m, 2H), 1.35 (d, J = 20.7 Hz, 6H). Five unobserved exchangeable protons.

Example 23 (R)-8-hydroxy-5-(1-hydroxy-2-(3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)methyl)phenethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

a) (R)-5-(1-(Third-butyldimethylmethylalkyloxy)-2-(3-((4-(2-isopropylthiazol-4-carbonyl)-1-) Oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)-8-hydroxyquinoline-2(1H)-one

(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-isopropylthiazole- A solution of 4-yl)methanone (Example 22, step c) (0.159 g) in EtOAc (EtOAc) (EtOAc) The resulting mixture was stirred at 20 ° C for 1 hour. The reaction mixture was treated with aq. sodium sulphate (10 mL) and EtOAc (EtOAc) The mixture was extracted with EtOAc EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (2mL) and added to (R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8-hydroxy-quinolin-2 (1H)-ketone (WO 2004106333) (0.12 g) and acetic acid (0.021 mL) in methanol (10 mL). The mixture was cooled in an ice-bath and EtOAc (EtOAc:EtOAc. Methanol was removed under reduced pressure and the residue was crystalljjjjjjjjjjjjjj The crude product was purified by flash chromatography on EtOAc (EtOAc m. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.127 g.

m/z760(M+H) ⁺ (APCI)

b) (R)-8-Hydroxy-5-(1-hydroxy-2-(3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diaza) Heterospiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

( R )-5-(1-(Third-butyldimethylmethylalkyloxy)-2-(3-((4-(2-isopropylthia)-4-carbonyl)-1-oxo) Hetero-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)-8-hydroxyquinoline-2(1H)-one (Example 23, a solution of step a) (0.127 g) in THF (4 mL) EtOAc (EtOAc) hour. The solvent was removed under reduced pressure and the crude material was purified by preparative HPLC (Sunfire ^(s) , gradient: 10 to 40% acetonitrile in 0.2% aqueous TFA). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.085 g.

m/z 646(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.16 (d, J = 36.5 Hz, 1H), 7.94 (s, 1H), 7.45-7.33 (m, 4H), 7.13 (d, J = 26.1 Hz, 1H), 6.99 (d, J = 24.2 Hz, 1H), 6.54 (d, J = 24.2 Hz, 1H), 5.38-5.32 (m, 1H), 4.29 (s, 2H), 3.70 (s, 4H), 3.64 (s, 2H), 3, 32-3.24 (m, 3H), 3.21-3.00 (m, 8H), 2.08-1.97 (m, 2H), 1.82-1.68 (m, 2H), 1.34 ( d, J = 25.7 Hz, 6H). Six unobserved exchangeable protons.

Example 24 (R)-4-hydroxy-7-(1-hydroxy-2-(3-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)ethoxy)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(3-(2,2-diethoxyethoxy)phenyl)ethanol

Cesium carbonate (8.84 g) was added to a solution of 3-(2-hydroxyethyl)phenol (2.5 g) and 2-bromo-1,1-diethoxyethane (3.74 g) in DMF (50 mL) The mixture formed was stirred at 90 C overnight and then allowed to cool. The mixture was poured into water and extracted three times with ethyl acetate. The combined extracts were washed with water and brine, then dried over anhydrous sodium sulfate. The yield was 3.0 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.21-7.16 (m, 1H), 6.82-6.72 (m, 3H), 4.80 (t, J = 5.3 Hz, 1H), 3.97 (d, J = 5.1 Hz, 2H), 3.81 (t, J = 6.5 Hz, 2H), 3.78-3.69 (m, 2H), 3.65-3.56 (m, 2H), 2.80 (t, J = 6.5 Hz, 2H), 1.21 (t, J) =6.9 Hz, 6H). An unobserved exchangeable proton.

b) 2-(3-(2-hydroxyethyl)phenoxy)acetaldehyde

Concentrated hydrochloric acid (1.5 mL) was added to 2-(3-(2,2-diethoxyethoxy)phenyl)ethanol (Example 24, step a) (0.256 g) in 1,4-dioxin In a solution of the alkane (3 mL), and the mixture formed was stirred at room temperature for 1.5 hours. The solution was then diluted with water and extracted twice with ethyl acetate. The combined extracts were washed with brine, dried over anhydrous magnesium The yield was 0.150 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ9.86 (s, 1H), 7.28-7.13 (m, 1H), 6.92-6.66 (m, 3H), 4.57 (s, 2H), 3.87 (t, J = 6.5 Hz, 2H), 2.86 (t, J = 6.5 Hz, 2H). An unobserved exchangeable proton.

c) (9-(2-(3-(2-hydroxyethyl)phenoxy)ethyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl (2-methylthiazol-4-yl)methanone

2-(3-(2-hydroxyethyl)phenoxy)acetaldehyde (Example 24, step b) (0.143 g) and (2-methylthiazol-4-yl)(1-oxa-4, A suspension of 9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 4, step h) (0.314 g) in methanol (10 mL) with acetic acid (0.045 (mL) and stirred at room temperature for 30 minutes. The mixture was cooled in ice-water, treated with sodium triethyl succinate (0.254 g) and stirred for 3 days to allow the ice bath to pass. The resulting solution was purified by flash chromatography on EtOAc (EtOAc:EtOAc:EtOAc The second purification was carried out by flash chromatography on EtOAc (EtOAc m.). The yield was 0.122 g.

m/z 446(M+H) ⁺ (APCI)

d) 2-(3-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl) Ethoxy)phenyl)acetaldehyde

(9-(2-(3-(2-hydroxyethyl)phenoxy)ethyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)( A solution of 2-methylthiazol-4-yl)methanone (Example 24, step c) (0.152 g) in EtOAc (EtOAc) 5 minutes. Dess-Martin periodinane (0.221 g) was added and the mixture was taken out from the cooling bath and stirred at room temperature for 25 minutes. The reaction was quenched by the addition of saturated aqueous sodium thiosulfate (5 mL), saturated sodium bicarbonate (5 mL) and ethyl acetate (5 mL) and the mixture was stirred for 10 min. The mixture was then extracted twice with ethyl acetate and the combined organic extracts were washed with brine. Acetic acid (0.1 mL) was added, and the EtOAc EtOAc m. The yield was 0.118 g.

m/z 444(M+H) ⁺ (APCI)

e) (R)-4-hydroxy-7-(1-hydroxy-2-(3-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)ethoxy)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

( R )-7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) A solution of 0.105 g) in MeOH (2 mL) was taken (EtOAc) (EtOAc) Subsequent addition of 2-(3-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl) A solution of ethoxy)phenyl)acetaldehyde (Example 24, step d) (0.117 g) in MeOH (3 mL), and the mixture was stirred at room temperature for 1 hour then cooled in ice-water and Treatment with sodium acetoxyborohydride (0.085 g). The mixture was stirred on ice for 1 hour and then more sodium tris-sodium hydride hydride (0.086 g). The mixture was stirred for 1 hour on ice, after which the mixture was added to sodium tris-sodium hydride hydride (0.169 g), and the mixture was stirred for 1 hour. More sodium triethoxysulfonium borohydride (0.506 g) was added, and the mixture was stirred overnight and allowed to warm slowly to room temperature. The next day, the mixture was again cooled in ice-water and treated with sodium <RTI ID=0.0></RTI></RTI><RTIID=0.0> The residue was dissolved in acetonitrile (3mL) and water (1.5mL) of the mixture and by purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile). The product containing fractions were concentrated in vacuo and co-evaporated twice from acetonitrile to give a colourless residue. The residue was triturated with EtOAc (EtOAc)EtOAc. The yield was 0.011 g.

m/z 654(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO) δ 11.68 (br s, 1H), 10.22 (br s, 1H), 8.00 (s, 1H), 7.29 (t, J = 7.9 Hz, 1H), 6.96- 6.84 (m, 4H), 6.77 (d, J = 8.5 Hz, 1H), 6.48 (br s, 1H), 4.93-4.85 (m, 1H), 4.36-4.27 (m, 2H), 3.83-3.61 (m , 6H), 3.60-3.51 (m, 2H), 3.50-3.42 (m, 2H), 3.24-3.02 (m, 6H), 3.02-2.84 (m, 2H), 2.69 (s, 3H), 2.15-2.06 (m, 2H), 1.88-1.67 (m, 2H). Three unobserved exchangeable protons.

Example 25 (R)-8-hydroxy-5-(1-hydroxy-2-(3-((4-(2-(trifluoromethyl)thiazole-4-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

a) 1-oxa-4,9-diazaspiro[5.5]undecane-4-yl(2-(trifluoromethyl)thiazol-4-yl)methanone trifluoroacetate

1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester hydrochloride (WuXi PharmaTech) (0.594 g) and 2-(trifluoromethyl)thiazole- A solution of 4-carboxylic acid (0.4 g) in DMF (10 mL) HATU (1.003 g) was added, and the mixture was stirred at 20 ° C for 2 hours. The mixture was partitioned between EtOAc and EtOAc. The crude product was purified by flash chromatography on silica gel eluting with 40% ethyl acetate in isohexane to yield the product as a free base. This material was dissolved in dichloromethane (40 mL) and was taken &lt;RTI ID=0.0&gt;&gt; Toluene (60 mL) was added and the solvent was evaporated under reduced pressure to give subtitle compound. The yield was 0.740 g.

m/z 336(M+H) ⁺ (APCI)

b) (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-(trifluoro) Methyl)thiazol-4-yl)methanone

2-(3-(Bromomethyl)phenyl)ethanol (Example 6, step a) (0.354 g) was added to 1-oxa-4,9-diazaspiro[5.5]undecane- 4-(2-(Trifluoromethyl)thiazol-4-yl)methanone trifluoroacetate (Example 25, Step a) (0.74 g) and triethylamine (0.689 mL) in acetonitrile (15 mL) The solution was stirred and the mixture was stirred at 20 ° C for 2 hours. The solvent was evaporated under reduced pressure and the~~~~~~ The crude product was purified by flash chromatography on silica gel using 1% triethylamine and 3% methanol in dichloromethane. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.640 g.

m/z 470(M+H) ⁺ (APCI)

c) (R)-5-(1-(Third-butyldimethylmethylalkyloxy)-2-(3-((4-(2-(trifluoromethyl)thiazole-4-carbonyl)) -1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethylamine y)ethyl)-8-hydroxyquinoline-2(1H)- ketone

(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-(trifluoromethyl) a solution of thiazol-4-yl)methanone (Example 25, step b) (0.211 g) in dichloromethane (10 mL) with trifluoroacetic acid (0.035 mL), followed by Dess-Martin periodinane Treatment (0.266 g) and the resulting mixture was stirred at 20 ° C for 1 hour. The reaction mixture was treated with aq. sodium sulphate (10 mL) and EtOAc (EtOAc) The mixture was extracted with EtOAc (EtOAc)EtOAc.EtOAc. The residue was dissolved in methanol (2mL) and added to (R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8-hydroxy-quinolin-2 (1H)-ketone (WO 2004106333) (0.150 g) and acetic acid (0.026 mL) in MeOH (10 mL). The mixture was cooled in an ice-bath and EtOAc (EtOAc:EtOAc. Methanol was removed under reduced pressure and the residue was crystalljjjjjjjjjjjjjj The crude product was purified by flash chromatography on EtOAc (EtOAc m. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.11 g.

m/z 786(M+H) ⁺ (APCI)

d) (R)-8-hydroxy-5-(1-hydroxy-2-(3-((4-(2-(trifluoromethyl)thiazol-4-carbonyl)-1-oxa-4,9 -diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

Triethylamine trihydrofluoride (0.030 mL) in methanol (1 mL) was added to ( R )-5-(1-( T -butyldimethylmethylalkyloxy)-2-(3) -((4-(2-(trifluoromethyl)thiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)benzene Ethylamino)ethyl)-8-hydroxyquinolin-2(1H)-one (Example 25, step c) (0.11 g) in THF (4 mL). Leave for 18 hours. The solvent was removed under reduced pressure and the crude product was purified by preparative HPLC (Sunfire ^(TM) , gradient: 10 to 40% acetonitrile in 0.2% aqueous TFA). The fractions containing the desired compound were evaporated to dryness to give crystals crystals.

m/z 672(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ 8.78 (s, 1H), 8.52 (s, 1H), 8.18 (d, J = 49.2Hz, 1H), 7.45-7.33 (m, 4H), 7.14 (d, J = 21.8 Hz, 1H), 7.00 (d, J = 19.8 Hz, 1H), 6.54 (d, J = 21.9 Hz, 1H), 5.39 - 5.33 (m, 1H), 4.29 (s, 2H) ), 3.75-3.63 (m, 4H), 3.59 (s, 2H), 3.28 (t, J = 8.1 Hz, 2H), 3.21-3.00 (m, 8H), 2.10 - 1.97 (m, 2H), 1.84- 1.67 (m, 2H). Five unobserved exchangeable protons.

Example 26 (R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(2-(trifluoromethyl)thiazole-4-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-(trifluoromethyl) a solution of thiazol-4-yl)methanone (Example 25, step b) (0.268 g) in dichloromethane (10 mL) with trifluoroacetic acid (0.044 mL), followed by Dess-Martin periodinane Treatment (0.339 g) and the resulting mixture was stirred at 20 ° C for 1 hour. The reaction mixture was treated with aq. sodium sulphate (10 mL) and EtOAc (EtOAc) The mixture was extracted with EtOAc (EtOAc)EtOAc.EtOAc. The residue was dissolved in methanol (2 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.15) and acetic acid (0.033 mL) in 0 ° C in methanol (10 mL). Sodium cyanoborohydride (0.054 g) was added, and the mixture was stirred at room temperature for 18 hr. Most of the methanol was evaporated under reduced pressure and the residue was partitioned between THF (50 mL), brine (10 mL) and sat. The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.13 g.

m/z 678(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.27 (s, 1H), 8.74 (s, 1H), 8.52 (s, 1H), 7.45-7.31 (m, 4H), 6.93 (d, J=41.1 Hz, 1H), 6.77 (d, J=36.9 Hz, 1H), 4.97-4.87 (m, 1H), 4.27 (s, 2H), 3.74-3.69 (m, 2H), 3.69-3.64 (m , 2H), 3.59 (s, 2H), 3.25 (t, J = 14.5 Hz, 2H), 3.19-2.99 (m, 8H), 2.09-1.97 (m, 2H), 1.84-1.69 (m, 2H). Four unobserved exchangeable protons.

Example 27 (R)-4-mercapto-7-(1-hydroxy-2-(2-(5-((4-(5-methylthiophene-2-carbonyl)-1-oxo-4,9-di) Azaspiro[5.5]undecane-9-yl)methyl)thiophen-3-yl)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetic acid salt

a) tert-butyldimethyl(2-(thien-3-yl)ethoxy)decane

Addition of tert-ylchlorodimethyl decane (3.88 g) to 2-(thiophen-3-yl)ethanol (3.00 g) and 1H-imidazole (4.78 g) in DMF (30 mL) Stir the solution. After 16 h the reaction was diluted with EtOAc EtOAc m. Purification by chromatography on silica gel, eluting with isohexane and then 1:5 ethyl acetate:hexanes The yield was 5.2 g.

_{^{1 H NMR (300MHz, CDCl 3}} ) δ7.23 (dd, J = 2.9,4.9Hz, 1H), 7.01-6.99 (m, 1H), 6.97 (dd, J = 1.3,4.9Hz, 1H), 3.80 ( t, J = 6.8 Hz, 2H), 2.85 (t, J = 6.8 Hz, 2H), 0.88 (s, 9H), 0, 01 (s, 6H).

b) 4-(2-(Third-butyldimethylsilyloxy)ethyl)thiophene-2-carbaldehyde with 3-(2-(t-butyldimethyl decyloxy)B a mixture of thiophene-2-carbaldehyde

Add n-butyllithium (1.6 M in hexane, 7 mL) dropwise to tris-butyldimethyl(2-(thiophen-3-yl)ethoxy)decane (Example 27, step a) (2.210 g) in a stirred solution of tetrahydrofuran (60 mL) cooled at -78 °C. After the addition, the reaction mixture was stirred in an ice bath for 1 h and then cooled to -78. N,N-dimethylformamide (9.00 g) was added dropwise over 5 min and the cooling bath was removed after another 10 min. After 1 h, the reaction mixture was partitioned between EtOAc EtOAc. Purification by chromatography on silica gel, 1:20 ethyl acetate: isohexane eluting generated by ¹ H NMR is 4- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethyl) thiophene-2 An oil of a 4:1 mixture of formaldehyde and 3-(2-(tert-butyldimethylsilyloxy)ethyl)thiophene-2-carbaldehyde. The yield is 1.3g.

4-(2-(Third-butyldimethylsilyloxy)ethyl)thiophene-2-carbaldehyde:

_{^{1 H NMR (400MHz, CDCl 3}} ) δ9.93 (d, J = 1.2Hz, 1H), 7.71 (d, J = 1.5Hz, 1H), 7.52 (s, 1H), 3.92-3.84 (m, 2H) , 2.91 (t, J = 6.5 Hz, 2H), 0.92 (s, 9H), 0.04 (s, 6H).

3-(2-( Third -butyldimethylsilyloxy)ethyl)thiophene-2-carbaldehyde: ¹ H NMR (400 MHz, CDCl ₃ ) δ 10.08 (s, 1H), 7.69 (d, J=5.0 Hz, 1H), 7.09 (d, J=5.0 Hz, 1H), 3.92-3.84 (m, 2H), 3.22 (t, J=6.5 Hz, 2H), 0.89 (s, 9H), -0.01 (s, 6H).

c) (9-((4-(2-(Third-butyldimethylsilyloxy)ethyl)thiophen-2-yl)methyl)-1-oxa-4,9-diaza Heterospiro[5.5]undecane-4-yl)(5-methylthien-2-yl)methanone

(5-Methylthiophen-2-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 9, Step b) (0.230g) was added to 4- (2- (third - butyldimethylsilyl silicon alkyloxy) ethyl) thiophene-2-carbaldehyde and 3- (2- (third - butyldimethyl 4:1 mixture of decyloxy)ethyl)thiophene-2-carboxaldehyde (Example 27, step b) (0.20 g) and AcOH (0.033 mL) in N -methyl-2-pyrrolidone (3 mL) In the stirred solution. After 5 min, sodium triethoxysulfonate borohydride (0.35 g) was added. After 16 h, water was added and the mixture was extracted with ethyl acetate. The ethyl acetate layer was washed three times with water and evaporated in vacuo. Purification by silica gel chromatography, eluting with 20:80:5 ethyl acetate: isohexane: triethylamine, and separating the two isomeric products to give a sub-title compound. The yield was 0.21 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.09 (d, J = 3.8 Hz, 1H), 6.86 (s, 1H), 6.75 (s, 1H), 6.69 (d, J = 3.8 Hz, 1H), 3.78 (t, J = 6.6 Hz, 2H), 3.76-3.69 (m, 4H), 3.64 (s, 2H), 3.56 (s, 2H), 2.76 (t, J = 7.1 Hz, 2H), 2.62-2.54 ( m, 2H), 2.50 (s, 3H), 2.41-2.32 (m, 2H), 1.90 - 1.82 (m, 2H), 1.63-1.52 (m, 2H), 0.88 (s, 9H), 0.01 (s, 6H).

d) (9-((4-(2-hydroxyethyl))thiophen-2-yl)methyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl (5-methylthien-2-yl)methanone

Tetrabutylammonium fluoride (1 M in tetrahydrofuran, 2 mL) was added to (9-((4-(2-(t-butyl dimethyl decyloxy)ethyl) thiophen-2-yl)) Methyl)-1-oxa-4,9-diazaspiro[5.5]undecethane-4-yl)(5-methylthien-2-yl)methanone (Example 27, step c) (0.65 g) in a solution in tetrahydrofuran (7 mL). After 1 h, the solution was evaporated in vacuo. Purification by silica gel chromatography eluting with 20:1 ethyl acetate: triethylamine afforded the subtitle compound. The yield was 0.45 g.

¹ H NMR (400 MHz, CDl _{3 3} ) δ 7.09 (d, J = 3.7 Hz, 1H), 6.92 (s, 1H), 6.77 (s, 1H), 6.69 (dd, J = 1.0, 3.7 Hz, 1H) , 3.83 (t, J = 6.4 Hz, 2H), 3.76-3.70 (m, 4H), 3.66 (s, 2H), 3.56 (s, 2H), 2.82 (t, J = 6.4 Hz, 2H), 2.62 2.55 (m, 2H), 2.50 (s, 3H), 2.43-2.33 (m, 2H), 1.91-1.82 (m, 2H), 1.65-1.50 (m, 2H). An unobserved exchangeable proton.

e) 2-(5-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl Thiophen-3-yl)acetaldehyde

Add Dess-Martin periodinane (0.35 g) to (9-((4-(2-hydroxyethyl))thiophen-2-yl)methyl)-1-oxa-4,9-diaza Spiro [5.5] undecane-4-yl)(5-methylthiophen-2-yl)methanone (Example 27, step d) (0.19 g) and trifluoroacetic acid (0.052 mL) in DCM (5 mL) In the stirred solution. After 1 h, ethyl acetate (30 mL) was added followed by a mixture of saturated sodium thiosulfate (5 mL) and saturated sodium bicarbonate (5 mL). The reaction mixture was thoroughly shaken and separated. The ethyl acetate solution was washed with saturated sodium bicarbonate solution, water and brine. Acetic acid (0.07 mL) was added, the solution was dried over sodium sulfate, filtered and evaporated in vacuo. The yield was 0.19 g. Use directly.

f) (R)-4-hydroxy-7-(1-hydroxy-2-(2-(5-((4-(5-methylthiophen-2-carbonyl)-1-oxo-4,9-) Diazaspiro[5.5]undecane-9-yl)methyl)thiophen-3-yl)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoro Acetate

Acetic acid (0.039 g) was added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.17 g) and 2-(5-((4-(5-methylthiophen-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5] To a stirred solution of hexanes (10 mL) in MeOH (10 mL). After 1 min, sodium cyanoborohydride (0.10 g) was added. After 3 h, the~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ The organic solution was dried over sodium sulfate, filtered and evaporated in vacuo. The solid was dissolved in methanol and purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in acetonitrile 10 to 40%) by. The fractions containing the product were combined and evaporated to dryness in vacuo. Acetonitrile was added and the solution was evaporated in vacuo and the procedure was repeated. Diethyl ether was added and the gum was triturated to give the title compound as a solid. The yield was 0.097 g.

m/z 629(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ11.67 (s, 1H), 10.23 (s, 1H), 9.82-9.67 (m, 1H), 8.93-8.67 (m, 2H), 7.46 (s, 1H ), 7.25-7.22 (m, 1H), 7.17 (s, 1H), 6.93 (d, J = 8.8 Hz, 1H), 6.77 (d, J = 8.8 Hz, 1H), 6.60-6.46 (m, 1H) , 6.51-6.46 (m, 1H), 4.92-4.86 (m, 1H), 4.68-4.51 (m, 2H), 3.74-2.88 (m, 16H), 2.46 (s, 3H), 2.15-2.04 (m, 2H), 1.69-1.56 (m, 2H).

Example 28 (R)-5-(2-(3-fluoro-5-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] eleven Cycloalkyl-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetate

2-(3-Fluoro-5-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl) A solution of methyl (phenyl) acetaldehyde (Example 19, step c) (0.2 g) in methanol (3 mL) was added to ( R )-5-(2-amino-1-( tri -butyl) A mixture of dimethyl decyloxy)ethyl)-8-hydroxyquinolin-2(1H)-one (WO 2004106333) (0.23 g) and acetic acid (0.027 mL) in methanol (3 mL). The resulting mixture was stirred for 5 min and then cooled to 0 °C. Sodium cyanoborohydride (0.044 g) was then added and the mixture was warmed to RT and stirred for 2 h. The solvent was evaporated in vacuo and purified by silica gel chromatography eluting with gradient: 95:5:0.5 to 89:10:1 DCM:methanol: The fractions containing the product were combined and evaporated in vacuo. The residue was dissolved in THF (3 mL)EtOAcEtOAcEtOAc The reaction was concentrated in vacuo. Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile). The title compound was obtained as a white solid. The yield was 0.021 g.

m/z 636(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.15 (d, J = 10.0 Hz, 1H), 7.90 (s, 1H), 7.29-7.17 (m, 3H), 7.13 (d, J = 8.2 Hz, 1H), 6.99 (d, J = 8.2 Hz, 1H), 6.55 (d, J = 9.7 Hz, 1H), 5.37-5.29 (m, 1H), 4.35-4.19 (m, 2H), 3.75- 3.59 (m, 6H), 3.34 - 2.98 (m, 10H), 2.67 (s, 3H), 2.06-1.93 (m, 2H), 1.83-1.69 (m, 2H). Six unobserved exchangeable protons.

Example 29 (R)-7-(2-(4-fluoro-3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5] eleven Cycloalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(4-fluoro-3-methylphenyl)acetic acid

2-(4-Fluoro-3-methylphenyl)acetonitrile (1 g) and sodium hydroxide (0.8 g) were combined in a mixture of methanol (10 mL) and water (3 mL). The resulting mixture was then heated under reflux overnight. The reaction was concentrated in vacuo and the residue was crystalljjjjjjj The aqueous phase was washed with EtOAc (EtOAc (EtOAc)EtOAc. The combined organic solution was washed with EtOAc (EtOAc m. The yield is 1.1g.

¹ H NMR (300 MHz, D _{6 -} DMSO) δ 12.31 (s, 1H), 7.20-6.99 (m, 3H), 3.52 (s, 2H), 2.21. (s, 3H).

b) (9-(2-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(5 -methylthiophen-2-yl)methanone

Benzoyl peroxide (0.058 g) was added to 2-(4-fluoro-3-methylphenyl)acetic acid (0.6 g) (Example 29, step a) and N -bromosuccinimide (0.7 g) ) in a mixture of DCM (10 mL). The resulting mixture was heated under reflux for 4 h. DCM (10 mL) and water (20 mL) were added and the organic phase was separated. The organic phase was washed with brine (20 mL) dried over sodium sulfate. The residue was redissolved in THF (10 mL) and cooled in ice. A solution of borane dimethyl sulfide (2M in THF, 4.46 mL) was added dropwise and the mixture was stirred for 1 h. Methanol (2 mL) was carefully added and once the bubbling ceased, the solvent was evaporated in vacuo. The residue was redissolved in acetonitrile (10 mL) and (5-methylthiophen-2-yl)(1-oxa-4,9-diazaspiro[5.5]undec-4-yl)methanone was added. Trifluoroacetate (Example 9, step b) (0.7 g) followed by triethylamine (1.49 mL). The resulting mixture was stirred overnight and then concentrated in vacuo. Purification by gel chromatography, eluting with a gradient of 99:1:0.1 to 97:3:0.3 DCM:methanol: '880' aqueous ammonia solution. The fractions containing the product are combined and evaporated to yield a foam of the sub-title compound. The yield was 0.46 g.

m/z 433(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO) δ 7.22 - 7.16 (m, 2H), 7.14 - 7.07 (m, 1H), 6.97 (dd, J = 10.0, 8.5 Hz, 1H), 6.80-6.76 (m) , 1H), 3.67-3.57 (m, 6H), 3.50-3.45 (m, 4H), 2.70 (t, J = 6.8 Hz, 2H), 2.46 (s, 3H), 2.41-2.35 (m, 4H), 1.76-1.67 (m, 2H), 1.55-1.45 (m, 2H). An unobserved exchangeable proton.

c) (R)-7-(2-(4-fluoro-3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

Add TFA (0.032 mL) to (9-(2-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 4-Base)(5-Methylthiophen-2-yl)methanone (Example 29, step b) (0.18 g) in DCM (5 mL) The mixture was stirred for 5 min, then Dess-Martin periodinane (0.27 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (25 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (25 mL). The combined organic solution was washed with brine, acidified with a few drops of acetic acid, dried over sodium sulfate, filtered and evaporated in vacuo. The residue was dissolved in methanol (5 mL) followed by acetic acid (0.012 mL) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.17 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.039 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 30% acetonitrile). The title compound was obtained as a white solid. The yield was 0.11 g.

m/z 641(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ7.47-7.35 (m, 2H), 7.29-7.17 (m, 2H), 6.93 (d, J = 8.2Hz, 1H), 6.82-6.75 ( m, 2H), 4.91 (dd, J=8.1, 5.3 Hz, 1H), 4.32-4.20 (m, 2H), 3.74-3.61 (m, 4H), 3.53 (s, 2H), 3.28-2.93 (m, 10H), 2.46 (s, 3H), 2.05-1.97 (m, 2H), 1.82-1.74 (m, 2H). Six unobserved exchangeable protons.

Example 30 (R)-5-(9-(3-(2-(2-hydroxy-2-(4-hydroxy-2-yloxy-2,3-dihydrobenzo[d]thiazole-7-yl)) Ethylamino)ethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-carbonyl)thiophene-2-carboxylic acid methyl ester di-trifluoroacetate

a) 4-(5-(Methoxycarbonyl)thiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester

1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester hydrochloride (WuXi PharmaTech) (0.6 g) and thiophene-2,5-dicarboxylic acid (1.7) g) The solution in DMF (10 mL) was taken (3. HATU (1.013 g) was added, and the mixture was stirred at 20 ° C for 2 hours. The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The residue was purified by flash chromatography on EtOAc (EtOAc) elute This material was dissolved in THF (30 mL), EtOAc (EtOAc) Methanol (20 mL) was added and the reaction was heated at 50 ° C for 30 min. The solvent was removed under reduced pressure and the residue was partitioned between ethyl acetate and brine. The organic layer was dried with sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on silica gel eluting with 40% ethyl acetate in isohexane. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.45 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.72 (d, J = 3.8 Hz, 1H), 7.40 (d, J = 3.8 Hz, 1H), 3.85 (s, 3H), 3.72-3.68 (m, 2H), 3.63-3.59 (m, 2H), 3.58-3.51 (m, 2H), 3.49 (s, 2H), 3.12-3.04 (m, 2H), 1.77-1.70 (m, 2H), 1.46 - 1.40 (m, 2H), 1.39 (s, 9H).

b) 5-(1-oxa-4,9-diazaspiro[5.5]undecane-4-carbonyl)thiophene-2-carboxylic acid methyl ester trifluoroacetate

Trifluoroacetic acid (10 mL) was added to 4-(5-(methoxycarbonyl)thiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9- To the solution of the third butyl formate (Example 30, step a) (0.45 g) in dichloromethane (40 mL), and the mixture was allowed to stand at 20 ° C for 1 hour. Toluene (50 mL) was added and the solvent was removed under reduced pressure to give subtitle compound. The yield was 0.46 g.

m/z 325(M+H) ⁺ (APCI)

c) 5-(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-carbonyl)thiophene-2- Methyl formate

5-(1-oxa-4,9-diazaspiro[5.5]undecane-4-carbonyl)thiophene-2-carboxylic acid methyl ester trifluoroacetate (Example 30, step b) (0.46 g And a solution of 2-(3-(bromomethyl)phenyl)ethanol (Example 6, Step a) (0.226 g) in EtOAc (10 mL) Stir at °C for 2 hours. The solvent was evaporated under reduced pressure. The organic layer was dried with sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on silica gel using 1% triethylamine and 3% methanol in dichloromethane. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.41 g.

m/z 459(M+H) ⁺ (APCI)

d) (R)-5-(9-(3-(2-(2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazole-7-) Ethylamino)ethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-carbonyl)thiophene-2-carboxylic acid methyl ester di-trifluoro Acetate

5-(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-carbonyl)thiophene-2-carboxylic acid A solution of the ester (Example 30, step c) (0.262 g) in dichloromethane <RTI ID=0.0>(</RTI><RTIID=0.0> The mixture was stirred at 20 ° C for 1 hour. The reaction mixture was treated with aq. sodium sulphate (10 mL) and EtOAc (EtOAc) The mixture was extracted with EtOAc (EtOAc)EtOAc.EtOAc. The residue was dissolved in methanol (2 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.15) and acetic acid (0.033 mL) in 0 ° C in methanol (10 mL). Sodium cyanoborohydride (0.054 g) was added, and the mixture was stirred at room temperature for 18 hr. Most of the methanol was evaporated under reduced pressure and the residue was partitioned between THF (50 mL), brine (10 mL) and sat. The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.175 g.

m/z 667(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.27 (s, 1H), 7.72 (d, J = 3.8 Hz, 1H), 7.44 - 7.33 (m, 5H), 6.93 (d, J = 8.5 Hz, 1H), 6.77 (d, J = 8.2 Hz, 1H), 4.95-4.89 (m, 1H), 4.30 (s, 2H), 3.85 (s, 3H), 3.74 - 3.69 (m, 2H), 3.66-3.61 (m, 2H), 3.53 (s, 2H), 3.28-2.98 (m, 10H), 2.09-1.99 (m, 2H), 1.77 (s, 2H). Five unobserved exchangeable protons.

Example 31 (R)-5-(9-(3-(2-(2-hydroxy-2-(8-hydroxy-2-oxo-l,2-dihydroquinolin-5-yl)ethylamino) Ethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-carbonyl)thiophene-2-carboxylic acid methyl ester di-trifluoroacetate

a) (R)-5-(9-(3-(2-(2-(tert-butyldimethyl)alkyloxy)-2-(8-hydroxy-2-yloxy-1, 2-Dihydroquinolin-5-yl)ethylamino)ethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-carbonyl)thiophene- Methyl 2-formate

5-(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-carbonyl)thiophene-2-carboxylic acid A solution of the ester (Example 30, step c) (0.137 g) in dichloromethane (10 mL) eluting with EtOAc (EtOAc) The mixture was stirred at 20 ° C for 1 hour. The reaction mixture was treated with aq. sodium sulphate (10 mL) and EtOAc (EtOAc) The mixture was extracted with EtOAc (EtOAc) EtOAc (EtOAc m. . The aldehyde was dissolved in methanol (2 mL) and added to ( R )-5-(2-amino-1-(tris-butyldimethylmethylalkyloxy)ethyl)-8-hydroxyquinoline-2 (1H)-ketone (WO 2004106333) (0.10 g) and acetic acid (0.017 mL) in methanol (10 mL). The mixture was cooled in an ice-bath and EtOAc (EtOAc)EtOAc. Methanol was removed under reduced pressure and the residue was crystalljjjjjjjjjjjjjj The crude product was purified by flash chromatography on EtOAc (EtOAc m.) The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.108 g.

m/z 775(M+H) ⁺ (APCI)

b) (R)-5-(9-(3-(2-(2-hydroxy-2-(8-hydroxy-2-oxoyl-1,2-dihydroquinolin-5-yl)ethyl) Amino)ethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-carbonyl)thiophene-2-carboxylic acid methyl ester di-trifluoroacetate

(R)-5-(9-(3-(2-(2-(T-butyl-dimethyl decyloxy))-2-(8-hydroxy-2-yloxy-1,2- Dihydroquinolin-5-yl)ethylamino)ethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-carbonyl)thiophene-2- Methyl formate (Example 31, step a) (0.108 g) was taken in THF (4 mL) EtOAc (EtOAc) 18 hours. The solvent was removed under reduced pressure and the crude product was purified by preparative HPLC (Sunfire ^(TM) , gradient: 10 to 40% acetonitrile in 0.2% aqueous TFA). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.055 g.

m/z 661(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.18 (d, J = 9.7 Hz, 1H), 7.72 (d, J = 3.8 Hz, 1H), 7.45-7.32 (m, 5H), 7.14 ( d, J = 8.2 Hz, 1H), 7.00 (d, J = 8.2 Hz, 1H), 6.54 (d, J = 9.7 Hz, 1H), 5.40 - 5.34 (m, 1H), 4.30 (s, 2H), 3.85 (s, 3H), 3.75-3.70 (m, 2H), 3.66-3.61 (m, 2H), 3.53 (s, 2H), 3.29 (t, J = 8.1 Hz, 2H), 3.22-3.01 (m, 8H), 2.10 - 1.99 (m, 2H), 1.78 (s, 2H). Six unobserved exchangeable protons.

Example 32 (R)-7-(2-(3,4-difluoro-5-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(3,4-difluorophenyl)ethanol

A solution of borane dimethyl sulfide (2M in THF, 18.30 mL) was carefully added to a solution of 2-(3,4-difluorophenyl)acetic acid (2.1 g) in THF (20 mL) EtOAc. The resulting mixture was allowed to warm to RT and stirred for 1 h. The reaction was cooled in an ice bath and methanol (5 mL) was added dropwise. The reaction was stirred until bubbling ceased and the solvent was evaporated in vacuo. Purify by gel chromatography and elute with isohexane to 4:1 isohexane:ethyl acetate. The fractions containing the product are combined and evaporated in vacuo to give the sub-title compound as a clear oil. The yield was 1.92 g.

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 7.14-7.00 (m, 2H), 6.98-6.90 (m, 1H), 3.85 (t, J = 6.4Hz, 2H), 2.82 (t, J = 6.4Hz, 2H ). An unobserved exchangeable proton.

b) 2,3-difluoro-5-(2-hydroxyethyl)benzaldehyde

2,2,6,6-Tetramethylpiperidine (6.15 mL) was added to a solution of n-butyllithium in hexane at -70 °C (1.6M, 22.8 mL). A solution of 2-(3,4-difluorophenyl)ethanol (Example 32, step a) (1.92 g) in THF (25 mL). THF (25 mL) was added and the mixture was stirred at -70 ° C for 6 h. DMF (4.7 mL) was then added and the mixture was stirred at -70 ° C for 1 h. The mixture was then allowed to warm to RT and stirred for 70 h. Aqueous HCl (2 M, 10 mL) was then added, then ethyl acetate (20 mL) and layered. The aqueous phase was extracted with ethyl acetate (2×20 mL). The combined organic solution was washed with brine (20 mL) dried over magnesium sulfate. Purified by silica gel chromatography and eluted with a gradient of 4:1 to 2:1 isohexane: diethyl ether. The fractions containing the product were combined and evaporated in vacuo to give a sub-title compound. The yield was 1.9 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 10.33 (s, 1H), 7.54-7.47 (m, 1H), 7.39-7.32 (m, 1H), 3.92-3.86 (m, 2H), 2.88 (t, J) =6.3HZ, 2H). An unobserved exchangeable proton.

c) (9-(2,3-Difluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl (5-methylthien-2-yl)methanone

(5-Methylthiophen-2-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 9, Step b) (0.53 g) was added to 2,3-difluoro-5-(2-hydroxyethyl)benzaldehyde (Example 32, step b) (0.25 g) and acetic acid (0.08 mL) in methanol (5 mL) In the solution. The reaction was stirred for 30 min and cooled in an ice bath. Sodium cyanoborohydride (0.13 g) was then added and the mixture was warmed to RT and stirred for 18 h. The reaction was concentrated in vacuo. Purification by gel chromatography, eluting with a gradient of 99:1:0.1 to 97:3:0.3 DCM:methanol: '880' aqueous ammonia solution. The fractions containing the product are combined and evaporated in vacuo to give a sub-title compound. The yield was 0.08 g.

m/z 451(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.18 (d, J = 3.6 Hz, 1H), 7.14-7.06 (m, 1H), 7.04-6.99 (m, 1H), 6.80-6.77 ( m,1H),4.34-4.29(m,1H), 3.68-3.57(m,6H),3.53-3.46(m,4H), 2.70(t,J=6.7Hz,2H), 2.47-2.34(m, 7H), 1.77-1.67 (m, 2H), 1.55-1.45 (m, 2H).

d) (R)-7-(2-(3,4-Difluoro-5-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetic acid salt

Add TFA (0.013 mL) to (9-(2,3-difluoro-5-(2-hydroxyethyl)benzyl)

(1)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(5-methylthien-2-yl)methanone (Example 32, step c) 0.077 g) in DCM (2 mL) in EtOAc. The mixture was stirred for 5 min, then Dess-Martin periodinane (0.11 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (1 mL), sat. sodium bicarbonate (1 mL) and ethyl acetate (5 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (25 mL). The combined organic solution was washed with brine, acidified with a few drops of acetic acid, dried over sodium sulfate, filtered and evaporated in vacuo. The residue was dissolved in methanol (5mL), followed by addition of acetic acid (0.005 mL) and (R) -7- (2- amino-1-hydroxyethyl) -4-hydroxybenzo [d] thiazol -2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.045 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.016 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 30% acetonitrile). The title compound was obtained as a white solid. The yield was 0.052 g.

m/z 659(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.27 (s, 1H), 7.46-7.37 (m, 1H), 7.28-7.23 (m, 1H), 7.20 (d, J = 3.6Hz, 1H), 6.93 (d, J = 8.2 Hz, 1H), 6.81-6.74 (m, 2H), 4.91 (dd, J = 8.1, 5.0 Hz, 1H), 4.28-4.19 (m, 2H), 3.74 - 3.62 (m, 4H), 3.53 (s, 2H), 3.25 (t, J = 7.9 Hz, 2H), 3.15-2.95 (m, 8H), 2.46 (s, 3H), 2.01-1.93 (m, 2H), 1.80-1.67 (m, 2H). Five unobserved exchangeable protons.

Example 33 (R)-4-hydroxy-7-(1-hydroxy-2-(3-((8-(5-methylthiophene-2-carbonyl)-5-oxa-2,8-diazaspiro[ 3.5]decane-2-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 1-Diphenylmethylazetidin-3-one

Triethylamine (24.7 mL) was added to a solution of 1-diphenylmethylazetidin-3-ol (5 g) in DMSO (25 mL). A solution of pyridine trisulfide (18 g) in DMSO (65 mL) was added dropwise and the mixture was stirred at 20 ° C for 90 min. The mixture was poured onto ice/water and extracted twice with ethyl acetate. EtOAc EtOAc. The crude product was purified by flash chromatography on silica gel using 2% triethylamine in isohexane as solvent. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 2.9 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.50-7.46 (m, 4H), 7.30 (t, J = 11.4 Hz, 4H), 7.24 - 7.19 (m, 2H), 4.59 (s, 1H), 4.00 ( s, 4H).

b) 1-Diphenylmethyl-3-(trimethyldecyloxy)azetidin-3-carbonitrile

Tetrabutylammonium cyanide (0.283 g) in dichloromethane (50 mL) was added dropwise to the 1-diphenylmethylazetidin-3-one at 20 ° C under nitrogen for 15 minutes. Example 33, step a) (2.5 g) and a stirred solution of trimethyldecane cyanide (2.82 mL) in dichloromethane (50 mL). The mixture was stirred at 20 ° C for 1 hour. The reaction mixture was washed with EtOAcq. The yield was 3.5 g. Used directly without purification.

c) 3-(Aminomethyl)-1-diphenylmethylazetidin-3-ol

a solution of 1-diphenylmethyl-3-(trimethyldecyloxy)azetidin-3-carbonitrile (Example 33, step b) (3.5 g) in THF (50 mL) Borane methyl sulfide complex treatment (2M in THF, 20.8 mL), and the resulting mixture was heated at 70 ° C for 1 hour under nitrogen. The mixture was cooled to room temperature and the reaction was quenched with EtOAc (EtOAc)EtOAc. The mixture was stirred at 20 ° C for 1 hour and then at 55 ° C for 1 hour. The mixture was cooled to room temperature and treated with tetrabutylammonium fluoride (1M in THF, 15.6 mL). The solvent was evaporated under reduced pressure and the residue was partitioned between ethyl acetate and brine. The organic layer was dried with sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on EtOAc. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 1.95 g.

m/z 269(M+H) ⁺ (APCI)

d) N-((1-Diphenylmethyl-3-hydroxyazetidin-3-yl)methyl)-2-chloroacetamide

Chloroacetyl chloride (0.846 mL) was added dropwise to 3-(aminomethyl)-1-diphenylmethylazetidin-3-ol in ethyl acetate (100 mL) over 30 min. Example 33, step c) (2.1 g) and potassium carbonate (3.03 g) in water (100 mL) were stirred vigorously at 0 °C. The mixture was stirred at 0&lt;0&gt;C for additional 30 min then EtOAc (EtOAc)EtOAc. The yield was 2.6 g.

m/z 345(M+H) ⁺ (APCI)

e) 2-Diphenylmethyl-5-oxa-2,8-diazaspiro[3.5]decane-7-one

N -((1-Diphenylmethyl-3-hydroxyazetidin-3-yl)methyl)-2-chloroacetamide (Example 33, step d) (2.6 g) in THF ( 50 mL) was added dropwise to a vigorously stirred solution of potassium t-butoxide (1 M in toluol, 15.08 mL) and THF (150 mL) at EtOAc. After the addition was completed, the mixture was stirred at 75 ° C for 10 minutes and then cooled to room temperature. The solvent was removed under reduced pressure and the residue was partitioned between ethyl acetate and brine. The organic layer was dried with sodium sulfate, filtered andEtOAc evaporated The yield was 2.05 g.

m/z309(M+H) ⁺ (APCI)

f) 2-Diphenylmethyl-5-oxa-2,8-diazaspiro[3.5]decane

Borane methyl sulfide complex (2M in THF, 10.7 mL) was added to 2-diphenylmethyl-5-oxa-2,8-diazaspiro[3.5]decane-7-one ( Example 33, step e) (2 g) in EtOAc (40 mL)EtOAc. The mixture was cooled to room temperature and treated dropwise with methanol (40mL), followed by N1, N2 - dimethyl-ethane-1,2-diamine processing (3.43g). The mixture was heated at 70 ° C for 6 hours. The solvent was removed under reduced pressure and the residue was partitioned between ethyl acetate and brine. The organic layer was dried with sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on silica gel using 1% triethylamine and 5% methanol in dichloromethane. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 1.35 g.

m/z 295(M+H) ⁺ (APCI)

g) (2-Diphenylmethyl-5-oxa-2,8-diazaspiro[3.5]decane-8-yl)(5-methylthien-2-yl)methanone

HATU (2.267 g) was added in one portion to 5-methylthiophene-2-carboxylic acid (0.652 g) and 2-diphenylmethyl-5-oxa-2,8-diazaspiro[3.5]decane (Examples) 33, Step f) (1.35 g) and triethylamine (1.917 mL) in DMF (20 mL) in EtOAc. The cooling bath was removed and the mixture was stirred at room temperature for 2 hours. The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The crude product was purified by flash chromatography on EtOAc. The pure fractions were evaporated to dryness to give the subtitle compound. The yield is 1.5g.

m/z 419(M+H) ⁺ (APCI)

h) (5-methylthien-2-yl)(5-oxa-2,8-diazaspiro[3.5]decane-8-yl)methanone hydrochloride

1-chloroethyl chloroformate (0.508 mL) was added dropwise to (2-diphenylmethyl-5-oxa-2,8-diazaspiro[3.5]decane-8-yl) (5-A) Thiophen-2-yl)methanone (Example 33, step g) (1.5 g) in acetonitrile (30 mL) in EtOAc. The mixture was then heated under reflux for 1 hour under nitrogen. The solvent was removed under reduced pressure and the residue was evaporated mjjjjjjj This solution was heated under nitrogen for 30 minutes under reflux. The solvent was removed under reduced pressure and EtOAcqqqqqqqqqqq The yield was 0.29 g.

m/z 253(M+H) ⁺ (APCI)

i) (2-(3-(2-hydroxyethyl)benzyl)-5-oxa-2,8-diazaspiro[3.5]decane-8-yl)(5-methylthiophene-2) Ketone

2-(3-(Bromomethyl)phenyl)ethanol (Example 6, step a) (0.281 g) was added to (5-methylthiophen-2-yl)(5-oxa-2,8- A solution of diazaspiro[3.5]nonane-8-yl)methanone hydrochloride (Example 33, step h) (0.29 g) and triethylamine (0.42 mL) in acetonitrile (15 mL) at 20 ° C The mixture formed and stirred at 20 ° C for 3 hours. The solvent was removed under reduced pressure and the residue was partitioned between ethyl acetate and brine. The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.32 g.

m/z 387(M+H) ⁺ (APCI)

j) (R)-4-hydroxy-7-(1-hydroxy-2-(3-((8-(5-methylthiophene-2-carbonyl)-5-oxa-2,8-diaza Spiro[3.5]decane-2-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

(2-(3-(2-hydroxyethyl)benzyl)-5-oxa-2,8-diazaspiro[3.5]decane-8-yl)(5-methylthiophen-2-yl) a solution of ketone (Example 33, step i) (0.16 g) in dichloromethane (10 mL) EtOAc (EtOAc) The resulting mixture was stirred at 20 ° C for 1 hour. The reaction mixture was treated with aq. sodium sulphate (10 mL) and EtOAc (EtOAc) The mixture was extracted with EtOAc (EtOAc)EtOAc.EtOAc. The residue was dissolved in methanol (2 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.15) and acetic acid (0.024 mL) in 0 ° C in methanol (10 mL). Sodium cyanoborohydride (0.039 g) was added, and the mixture was stirred at room temperature for 18 hr. Most of the methanol was evaporated under reduced pressure and the residue was partitioned between THF (50 mL), brine (10 mL) and sat. The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.13 g.

m/z 595(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.26 (s, 1H), 7.42 - 7.29 (m, 4H), 7.24 (d, J = 3.6 Hz, 1H), 6.93 (d, J = 8.5 Hz, 1H), 6.82-6.75 (m, 2H), 4.95-4.88 (m, 1H), 4.36 (s, 2H), 4.09-3.97 (m, 4H), 3.87 (s, 2H), 3.73-3.69 (m, 2H), 3.66-3.62 (m, 2H), 3.24 (t, J = 8.1 Hz, 2H), 3.15-2.97 (m, 4H), 2.46 (s, 3H). Five unobserved exchangeable protons.

Example 34 (R)-7-(2-(2,6-Difluoro-3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (9-(2,4-Difluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl (5-methylthien-2-yl)methanone

Dibenzoyl peroxide (0.03 g) was added to a mixture of NBS (0.53 g) and 2-(2,6-difluoro-3-methylphenyl)acetic acid (0.5 g) in DCM (10 mL) . The reaction was heated at reflux for 4 h. DCM (10 mL) and water (20 mL) were added and the organic phase was separated. The organic phase was washed with brine (20 mL) dried over sodium sulfate. The residue was redissolved in THF (10 mL) and cooled in ice. A solution of borane dimethyl sulfide complex (2M in THF, 4 mL) was added dropwise and the mixture was stirred for 1 h. Methanol (2 mL) was added dropwise with caution and the solvent was evaporated once bubbling ceased. The residue was redissolved in acetonitrile (10 mL) and (5-methylthiophen-2-yl)(1-oxa-4,9-diazaspiro[5.5]undec-4-yl)methanone was added. Trifluoroacetate (Example 9, step b) (0.8 g) followed by triethylamine (1.12 mL). The resulting mixture was stirred overnight, evaporated and purified by EtOAc EtOAc (EtOAc) The yield was 0.37 g.

m/z 451(M+H) ⁺ (APCI)

b) 2-(2,6-Difluoro-3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane -9-yl)methyl)phenyl)acetaldehyde

Add TFA (0.05 mL) to (9-(2,4-difluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] eleven Carboxy-4-yl)(5-methylthiophen-2-yl)methanone (Example 34, step a) (0.27 g) in DCM (5 mL) in EtOAc. 5min. Dess-Martin periodinane (0.38 g) was then added and the mixture was stirred at RT for 45 min. Saturated sodium thiosulfate solution (5 mL), saturated aqueous sodium bicarbonate (5 mL) and ethyl acetate (20 mL) were then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organics were washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The yield was 0.27 g.

m/z449(M+H) ⁺ (APCI)

c) (R)-7-(2-(2,6-Difluoro-3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetic acid salt

2-(2,6-Difluoro-3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -Methyl)phenyl)acetaldehyde (Example 34, step b) (0.135 g) in methanol (3 mL) was added to (R)-7-(2-amino-1-hydroxyethyl) -4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.11 g) and acetic acid (0.015 mL) in methanol (0.5 mL) In the mixture. The resulting mixture was stirred for 5 min and then cooled to 0 °C. Sodium cyanoborohydride (0.025 g) was then added and the mixture was warmed to RT and stirred for 2 h. The reaction was concentrated and purified by silica gel chromatography eluting with a gradient of 95:5:0.5 to 89:10:1 DCM:MeOH: ' The combined product-containing fractions were dissolved, evaporated and purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile) by means of a white solid of the title compound. The yield is 0.1 g.

m/z 659(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.27 (s, 1H), 7.64 - 7.50 (m, 1H), 7.26-7.12 (m, 2H), 6.93 (d, J = 8.2 Hz, 1H), 6.84-6.69 (m, 2H), 4.91 (dd, J = 8.5, 4.9 Hz, 1H), 4.30 (s, 2H), 3.77-3.47 (m, 6H), 3.29-2.99 (m, 10H) , 2.46 (s, 3H), 2.10 - 1.90 (m, 2H), 1.88-1.68 (m, 2H). Five unobserved exchangeable protons.

Example 35 (R)-5-(2-(2,6-difluoro-3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetate

2-(2,6-Difluoro-3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -Methyl)phenyl)acetaldehyde (Example 34, step b) (0.135 g) in methanol (3 mL) was added to ( R )-5-(2-amino-1-( 3) a mixture of -butyldimethylmethylalkyloxy)ethyl)-8-hydroxyquinolin-2(1H)-one (WO 2004106333) (0.12 g) and acetic acid (0.017 mL) in methanol (3 mL) . The resulting mixture was stirred for 5 min and then cooled to 0 °C. Sodium cyanoborohydride (0.028 g) was then added and the mixture was warmed to RT and stirred for 2 h. The reaction was concentrated and the residue was purified using EtOAc EtOAc EtOAc:EtOAc:EtOAc The fractions containing the product were combined, evaporated and evaporated, mjjjjjjjjjjjjjjj The reaction was concentrated and the residue was purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile) to borrow. The title compound was obtained as a white solid. The yield was 0.12 g.

m/z 653(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.18 (d, J = 9.7 Hz, 1H), 7.64 - 7.51 (m, 1H), 7.22-7.10 (m, 3H), 7.00 (d, J = 8.2 Hz, 1H), 6.80 (d, J = 3.3 Hz, 1H), 6.54 (d, J = 10.0 Hz, 1H), 5.36 (dd, J = 8.7, 4.1 Hz, 1H), 4.34 (s, 2H), 3.76-3.46 (m, 6H), 3.33-3.03 (m, 10H), 2.46 (s, 3H), 2.09-1.95 (m, 2H), 1.87-1.66 (m, 2H). Six unobserved exchangeable protons.

Example 36 (R)-4-hydroxy-7-(1-hydroxy-2-(2-(5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxo-4,9-di) Azaspiro[5.5]undecane-9-yl)methyl)thiophen-3-yl)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetic acid salt

a) (9-((4-(2-(Third-butyldimethylsilyloxy)ethyl)thiophen-2-yl)methyl)-1-oxa-4,9-diaza Heterospiro[5.5]undecane-4-yl)(2-isopropylthiazol-4-yl)methanone

(2-Isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (0.50g) was added to 4- (2- (third - butyldimethylsilyl silicon alkyloxy) ethyl) thiophene-2-carbaldehyde and 3- (2- (third - butyldimethyl 4:1 mixture of decyloxy)ethyl)thiophene-2-carboxaldehyde (Example 27, step b) (0.40 g) and AcOH (0.085 mL) in N -methyl-2-pyrrolidone (6 mL) In the stirred solution. After 5 min, sodium triethoxysulfonate hydride (0.48 g) was added. After 16 h, water was added and the mixture was extracted with ethyl acetate. The ethyl acetate layer was washed three times with water and evaporated in vacuo. Purification by silica gel chromatography, eluting with ethyl acetate: isohexane: triethylamine 20: 80:5, and separating the two isomeric products to give a sub-title compound. The yield was 0.27 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.81 (s, 1H), 6.86 (s, 1H), 6.75 (s, 1H), 3.99-3.91 (m, 1H), 3.88-3.82 (m, 1H), 3.81-3.72 (m, 5H), 3.69-3.58 (m, 3H), 3.37-3.25 (m, 1H), 2.76 (t, J = 7.1 Hz, 2H), 2.59-2.44 (m, 3H), 2.41 2.29 (m, 1H), 1.89-1.81 (m, 2H), 1.78-1.67 (m, 1H), 1.56-1.50 (m, 1H) 1.46-1.34 (m, 6H), 0.87 (s, 9H), 0.00 (s, 6H).

b) (9-((4-(2-hydroxyethyl))thiophen-2-yl)methyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl (2-isopropylthiazol-4-yl)methanone

Tetrabutylammonium fluoride (in tetrahydrofuran 1M, 2mL) was added to (9 - ((4- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethyl) thiophen-2-yl) Methyl)-1-oxa-4,9-diazaspiro[5.5]undecyl-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 36, Step a (0.27 g) in a solution in tetrahydrofuran (4 mL). After 1 h, the solution was evaporated in vacuo. Purification by silica gel chromatography, eluting with ethyl acetate: triethylamine 20:1 affords the subtitle compound. The yield was 0.18 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.81 (s, 1H), 6.91 (s, 1H), 6.77 (s, 1H), 4.00-3.90 (m, 1H), 3.87-3.80 (m, 3H), 3.79-3.72 (m, 3H), 3.71-3.60 (m, 3H), 3.36-3.25 (m, 1H), 2.82 (t, J = 6.6 Hz, 3H), 2.61-2.42 (m, 3H), 2.42- 2.30 (m, 1H), 1.90 - 1.82 (m, 2H), 1.78-1.67 (m, 1H), 1.67-1.58 (m, 1H), 1.44-1.36 (m, 6H).

c) 2-(5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)) Thiophen-3-yl)acetaldehyde

Add Dess-Martin periodine (0.25g) to (9-((4-(2-hydroxyethyl))thiophen-2-yl)methyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 36, step b) (0.17 g) and trifluoroacetic acid (0.07 mL) in DCM ( In a stirred solution in 5 mL). After 1 h, ethyl acetate (30 mL) was added followed by a mixture of saturated sodium thiosulfate (5 mL) and saturated sodium bicarbonate (5 mL). The reaction mixture was thoroughly shaken and separated. The ethyl acetate solution was washed with saturated sodium bicarbonate solution, water and brine. Acetic acid (0.07 mL) was added, the solution was dried over sodium sulfate, filtered and evaporated in vacuo. The yield was 0.19 g. Use directly.

d) (R)-4-hydroxy-7-(1-hydroxy-2-(2-(5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxo-4,9) -diazaspiro[5.5]undecane-9-yl)methyl)thiophen-3-yl)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-three Fluoroacetate

Acetic acid (0.037 mL) was added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.170 g) and 2-(5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] A solution of the undecane-9-yl)methyl)thiophen-3-yl)acetaldehyde (Example 36, step c) (0.170 g) in methanol (8 mL). After 1 min, sodium cyanoborohydride (0.08 g) was added. After 3 h, the~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ The organic solution was dried over sodium sulfate, filtered and evaporated in vacuo. The solid was dissolved in methanol and purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in acetonitrile 10 to 40%) by. The fractions containing the pure product were combined and evaporated to dryness. The title compound was obtained as a white solid. The yield was 0.026 g.

m/z 658(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ 11.67 (s, 1H), 10.24 (s, 1H), 10.09-9.78 (m, 1H), 8.91-8.68 (m, 2H), 8.03 (s, 1H) , 7.46 (s, 1H), 7.17 (s, 1H), 6.93 (d, J = 8.9 Hz, 1H), 6.77 (d, J = 8.9 Hz, 1H), 6.51-6.46 (m, 1H), 4.93 4.85 (m, 1H), 4.67-4.51 (m, 2H), 3.85-2.88 (m, 17H), 2.14-2.05 (m, 2H), 1.82-1.58 (m, 2H), 1.34 (d, J = 6.8) Hz, 6H).

Example 37 (R)-5-(2-(3-((2,2-difluoro-4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetate

a) 4-((2-bromo-2,2-difluoroacetamido)methyl)-4-hydroxypiperidine-1-carboxylic acid tert-butyl ester

a solution of 4-(aminomethyl)-4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (1.702 g) in DMF (20 mL) at 20 ° C with 2-bromo-2,2-difluoroacetic acid Ester treatment (1.5 g) and the mixture was stirred at 20 ° C for 90 min under nitrogen. The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The yield is 2.5g.

m/z 385/387(MH) ^- (APCI)

b) 2,2-difluoro-3-indolyl-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester

3-((2-Bromo-2,2-difluoroacetamido)methyl)-4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (Example 37, step a) (3.2 g) in THF The solution in (40 mL) was added dropwise to a stirred solution of potassium tributoxide (1M in toluol, 16.53 mL) and THF (60 mL). At the end of the addition, the mixture was heated for another 10 minutes and then cooled to room temperature. The mixture was partitioned between EtOAc and EtOAcq. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.55 g.

m/z 305(MH) ^- (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO) δ 8.98 (s, 1H), 3.74 (d, J = 13.3 Hz, 2H), 3.43 (d, J = 2.8 Hz, 2H), 3.05 (s, 2H) , 1.81-1.73 (m, 2H), 1.70-1.61 (m, 2H), 1.40 (s, 9H).

c) 2,2-difluoro-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester

Borane methyl sulfide complex (2M in THF, 2.69 mL) was added dropwise to 2,2-difluoro-3-oxo-1-oxa-4,9-diazaspiro[5.5] A solution of undecane-9-carboxylic acid tert-butyl ester (Example 37, step b) (0.55 g) in THF (15 mL). The reaction mixture was heated at 55 ° C for 25 minutes under nitrogen and then cooled to room temperature. The mixture was quenched by the careful addition of methanol dropwise (5 mL). Subsequently, N1,N2-dimethylethane-1,2-diamine (0.633 g) was added and the mixture was heated at 70 ° C for 40 minutes. The solvent was removed under reduced pressure and the residue was purified by flash chromatography eluting with EtOAc The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.32 g. Use directly.

d) 2,2-difluoro-4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tertidine ester

HATU (0.541 g) was added in one portion to 2,2-difluoro-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester (Example 37, step c (0.32g) and a stirred solution of 5-methylthiophene-2-carboxylic acid (0.171g) and triethylamine (0.458mL) in DMF (20mL). The mixture was then stirred at 20 ° C for 7 hours. The mixture was partitioned between EtOAc and EtOAc. The crude product was purified by flash chromatography on flash chromatography eluting with 0.5 to 5% methanol in dichloromethane. The pure fractions were evaporated to dryness to give the subtitle compound. The yield is 0.4g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.29 (d, J = 3.6 Hz, 1H), 6.84 (dd, J = 3.6, 1.0 Hz, 1H), 4.08 (t, J = 8.8 Hz, 2H), 3.79 (s, 2H), 3.65-3.58 (m, 2H), 3.21-3.11 (m, 2H), 1.81-1.73 (m, 2H), 1.66-1.56 (m, 2H), 1.39 (s, 9H) + 3 unobserved H (methyl) (under solvent).

e) (2,2-difluoro-1-oxa-4,9-diazaspiro[5.5]undecethane-4-yl)(5-methylthiophen-2-yl)methanone trifluoro Acetate

Adding trifluoroacetic acid (5 mL) to 2,2-difluoro-4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane 9-carboxylic acid tert-butyl ester (Example 37, step d) (0.4 g) in dichloromethane (20 mL) in 20 ° C. The solution was allowed to stand at 20 ° C for 25 minutes. Toluene (40 mL) was added and the solvent was removed under reduced pressure. The residue was distilled twice with acetonitrile. The resulting gel was triturated with diethyl ether to give the sub-title compound. The yield was 0.39 g.

m/z 317(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ8.53 (s, 2H), 7.31 (d, J = 3.8Hz, 1H), 6.88-6.85 (m, 1H), 4.14 (t, J = 8.8 Hz, 2H), 3.84 (s, 2H), 3.27-3.21 (m, 2H), 3.13-3.04 (m, 2H), 2.05-1.98 (m, 2H), 1.91-1.81 (m, 2H). No methyl protons were observed (under solvent).

f) (2,2-Difluoro-9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl (5-methylthien-2-yl)methanone

2-(3-(Bromomethyl)phenyl)ethanol (Example 6, step a) (0.095 g) was added to (2,2-difluoro-1-oxa-4,9-diazaspiro) [5.5] undecane-4-yl)(5-methylthiophen-2-yl)methanone trifluoroacetate (Example 37, step e) (0.19 g) and triethylamine (0.185 mL) The solution was stirred at 20 ° C for 3 hours in a solution of acetonitrile (10 mL). The solvent was evaporated under reduced pressure and EtOAc evaporated m. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.188 g.

m/z 451(M+H) ⁺ (APCI)

g) (R)-5-(1-(Third-butyldimethylmethylalkyloxy)-2-(3-((2,2-difluoro-4-(5-methylthiophene-2) -carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)-8-hydroxyquinoline-2 ( 1H)-ketone

(2,2-Difluoro-9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)( A solution of 5-methylthiophen-2-yl)methanone (Example 37, step f) (0.188 g) in dichloromethane (10 mL) eluting with trifluoroacetic acid (0.032 mL) The mixture was treated with iodosane (0.248 g) and the resulting mixture was stirred at 20 ° C for 1 hour. The reaction mixture was treated with aq. sodium sulphate (10 mL) and EtOAc (EtOAc) The mixture was extracted with EtOAc (EtOAc)EtOAc.EtOAc. The residue was dissolved in methanol (2mL) and added to (R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8-hydroxy-quinolin-2 (1H)-ketone (WO 2004106333) (0.14 g) and acetic acid (0.024 mL) in methanol (10 mL). The mixture was cooled in an ice-bath and EtOAc (EtOAc:EtOAc. Methanol was removed under reduced pressure and the residue was crystalljjjjjjjjjjjjjj The crude product was purified by flash chromatography on EtOAc (EtOAc) eluting The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.114 g.

m/z 767(M+H) ⁺ (APCI)

h)(R)-5-(2-(3-((2,2-difluoro-4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetate

Triethylamine trihydrofluoride (0.03 mL) in methanol (1 mL) was added to ( R )-5-(1-( T -butyldimethylmethylalkyloxy)-2-(3) -((2,2-difluoro-4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)- a solution of phenylethylamino)ethyl)-8-hydroxyquinolin-2(1H)-one (Example 37, step g) (0.114 g) in THF (4 mL) Leave at 20 ° C for 18 hours. The solvent was removed, the crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in acetonitrile 10 to 40%) by under reduced pressure. The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.080 g.

m/z 653(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.16 (d, J = 9.7 Hz, 1H), 7.46-7.26 (m, 5H), 7.14 (d, J = 8.2 Hz, 1H), 6.99 (d, J = 8.2 Hz, 1H), 6.86-6.83 (m, 1H), 6.57-6.52 (m, 1H), 5.37-5.31 (m, 1H), 4.25 (s, 2H), 4.13 (t, J = 8.8 Hz, 2H), 3.82 (s, 2H), 3.30-2.99 (m, 10H), 2.10 - 1.88 (m, 4H). Six unobserved exchangeable protons. No methyl protons were observed (under solvent).

Example 38 (R)-4-hydroxy-7-(1-hydroxy-2-(2-(3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diaza) Heterospiro[5.5]undecane-9-yl)methyl)phenoxy)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (3-(2,2-diethoxyethoxy)phenyl)methanol

Cesium carbonate (7.87 g) was added to a solution of 3-(hydroxymethyl)phenol (2.5 g) and 2-bromo-1,1-diethoxyethane (3.97 g) in DMF (40 mL). The resulting mixture was heated at 90 ° C for 18 hours. The mixture was cooled to rt. EtOAc (EtOAc m. The crude product was purified by flash chromatography on silica gel eluting with 30% ethyl acetate in isohexane. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 1.9 g.

¹ H NMR (400 MHz, D _{6 -} DMSO) δ 7.21. (t, J = 7.9 Hz, 1H), 6.91-6.86 (m, 2H), 6.82-6.78 (m, 1H), 5.14 (t, J = 5.9 Hz, 1H), 4.79 (t, J = 5.1 Hz, 1H), 4.46 (d, J = 5.9 Hz, 2H), 3.93 (d, J = 5.1 Hz, 2H), 3.71-3.63 (m, 2H), 3.61-3.52 (m, 2H), 1.14 (t, J = 7.0 Hz, 6H).

b) (9-(3-(2,2-diethoxyethoxy)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl) (5-methylthien-2-yl)methanone

Methanesulfonium chloride (0.1 mL) was added dropwise to (3-(2,2-diethoxyethoxy)phenyl)methanol (Example 38, Step a) (0.307 g) and triethylamine ( 0.178 mL) stirred solution at 0 ° C in dichloromethane (30 mL). The resulting mixture was stirred at 20 ° C for 1 hour. The mixture was washed with water and the org. The residue was dissolved in acetonitrile (30 mL) eluted with triethylamine (1 mL), followed by (5-methylthiophen-2-yl)(1-oxa-4,9-diazaspiro[5.5] Monooxan-4-yl)methanone trifluoroacetate (Example 9, step b) (0.42 g). The mixture was stirred at 20 ° C for 2 hours. The solvent was removed under reduced pressure and the residue was crystalljjjjjjjjjjjjjj The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.45 g.

m/z 503(M+H) ⁺ (APCI)

c) 2-(3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Undecyl-9-yl)methyl)phenoxy)acetaldehyde

(9-(3-(2,2-Diethoxyethoxy)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(5 A solution of -methylthiophen-2-yl)methanone (Example 38, step b) (0.45 g) in a mixture of acetic acid (25 mL) and water (25 mL). Most of the solvent was removed under reduced pressure and the residue was partitioned between ethyl acetate and excess saturated sodium bicarbonate. The organic layer was dried with sodium sulfate and filtered andEtOAcEtOAcEtOAc The yield was 0.38 g. Use directly.

d) (R)-4-hydroxy-7-(1-hydroxy-2-(2-(3-((4-(5-methylthiophen-2-yl))-1-oxo-4,9- Diazaspiro[5.5]undecane-9-yl)methyl)phenoxy)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

Sodium cyanoborohydride (0.084 g) was added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.303 g) and 2-(3-((4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)methyl)phenoxy)acetaldehyde (Example 38, step c) (0.38 g) and acetic acid (0.051 mL) in methanol (12 mL) stirred at 20 ° C In solution. The reaction mixture was stirred at 20 ° C for 5 hours. The reaction mixture was evaporated to dryness (3 mL) eluting The aqueous layer was treated with saturated sodium bicarbonate solution (3 mL). The aqueous mixture was then treated with solid sodium chloride to give a saturated solution which was extracted with THF (20mL). The THF layer was dried over sodium sulfate, filtered and evaporated. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield is 0.1 g.

m/z 639(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.28 (s, 1H), 7.43-7.36 (m, 1H), 7.21 (s, 1H), 7.14 (s, 2H), 7.07 (d, J=8.5 Hz, 1H), 6.95 (d, J=8.2 Hz, 1H), 6.82-6.76 (m, 2H), 5.00-4.91 (m, 1H), 4.36-4.28 (m, 4H), 3.73-3.69 (m, 2H), 3.68-3.63 (m, 2H), 3.54 (s, 2H), 3.45 (s, 2H), 3.25-3.04 (m, 6H), 2.46 (s, 3H), 2.09-1.98 (m , 2H), 1.87-1.72 (m, 2H). Five unobserved exchangeable protons.

Example 39 7-((1R)-2-(2-fluoro-2-(3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenyl)ethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoro Acetate

a) ethyl 2-(3-bromophenyl)-2-fluoroacetate

2- (3-bromophenyl) acetate (1.00 g of) and the third - Silane-butyldimethylsilyl chloride (0.749 g) in THF (4mL) was cooled in the atmosphere at a nitrogen to -78 ℃, to Treatment with lithium bis(trimethyldecyl) aminide (1 M in THF, 4.6 mL) was added dropwise over 10 min. The solution was stirred at -78 ° C for 5 minutes, then taken out of the cooling bath and allowed to warm to room temperature over 30 minutes. The solution was concentrated in vacuo and the residue was suspended in isohexane and filtered to remove precipitated lithium chloride. The filtrate was concentrated in vacuo to give EtOAc ⁽ EtOAc ⁾ (EtOAc ⁾ 3 x 1 mL) was added. The resulting mixture was stirred for 1.5 hours, then concentrated on ruthenium dioxide and dried over EtOAc (EtOAc) eluting with EtOAc. oil. The yield was 0.884 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.63 (s, 1H), 7.53 (d, J = 7.9 Hz, 1H), 7.28 (t, J = 8.0 Hz, 2H), 5.73 (d, J = 47.7 Hz) , 1H), 4.33-4.19 (m, 2H), 1.28 (t, J = 7.1 Hz, 3H).

b) 2-(3-bromophenyl)-2-fluoroethanol

Lithium aluminum hydride (1 M in THF, 13.0 mL) was added portion EtOAc (EtOAc m. 35 mL) in a pre-cooled solution in ice-water. The resulting mixture was stirred in ice-water for 30 min then quenched (5 mL) by carefully adding methanol in portions over 30 min. The mixture was poured into 2M aqueous HCl and extracted with EtOAc EtOAc. The combined organic extracts were washed with EtOAc EtOAc EtOAc m. The yield was 1.39 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.53 - 7.46 (m, 2H), 7.30 - 7.24 (m, 2H), 5.53 (ddd, J = 48.2, 7.2, 3.3 Hz, 1H), 3.97 - 3.77 (m) , 2H), 1.95 (dd, J = 8.2, 5.1 HZ, 1H).

c) (2-(3-bromophenyl)-2-fluoroethoxy)(tri-butyl)dimethyl decane

A solution of 2-(3-bromophenyl)-2-fluoroethanol (Example 39, step b) (1.38 g) and imidazole (1.29 g) in DMF (14 mL) Treated with butyldimethylchloromethane (1.049 g), taken from a cooling bath and stirred overnight at room temperature. The solution was poured into water and extracted twice with diethyl ether. The combined organic extracts were washed twice with water and once with brine, then dried over anhydrous magnesium sulfate and purified by flash chromatography over EtOAc (EtOAc) Colorless oil. The yield was 1.94 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 7.51-7.49 (m, 1H), 7.46 (dt, J = 7.0,2.0Hz, 1H), 7.28-7.21 (m, 2H), 5.43 (ddd, J = 47.5, 6.5, 3.8 Hz, 1H), 3.95-3.76 (m, 2H), 0.88 (s, 9H), 0.03 (d, J = 4.6 Hz, 6H).

d) 3-(2-(Third-butyldimethylsilyloxy)-1-fluoroethyl)benzaldehyde

(2- (3-bromophenyl) -2-fluoroethoxy) (tertiary-butyl) - dimethyl Silane (Example 39, step c) (1.8g) in (36mL) in THF to a solution of It was cooled to -78 ° C under a nitrogen atmosphere and treated with butyl lithium (1.8M in hexane, 3.3 mL) dropwise over 5 min. The solution was stirred at -78 ° C for 30 minutes, treated with N , N -dimethylformamide (0.63 mL), stirred at -78 ° C for an additional 30 minutes, then taken out from the cooling bath and allowed to warm to room temperature over 140 minutes. . The solution was quenched by the addition of 10% aqueous ammonium chloride solution, and the resulting mixture was extracted twice with diethyl ether. The combined organics were washed with EtOAc (EtOAc m. The yield was 1.26 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 10.04 (s, 1H), 7.88-7.84 (m, 2H), 7.63 (d, J = 7.4Hz, 1H), 7.56 (t, J = 7.4Hz, 1H), 5.55 (ddd, J = 47.3, 6.2, 4.0 Hz, 1H), 4.00 - 3.83 (m, 2H), 0.87 (s, 9H), 0.02 (d, J = 3.9 Hz, 6H).

e) (9-(3-(2-(Terve-butyldimethylsilyloxy)-1-fluoroethyl)benzyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-4-yl)(2-isopropylthiazol-4-yl)methanone

3- (2- (Third - butyldimethylsilyl silicon alkyloxy) -1-fluoroethyl) benzaldehyde (example 39, step d) (4mL) in the solution (0.329 g) in MeOH acetic acid Treatment (0.055 mL) followed by (2-isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoride Acetate treatment (Example 22, step b) (0.408 g) was stirred at room temperature for 5 min. The solution was cooled in an ice-water-bath, EtOAc (EtOAc m. The solution was cooled back in ice-water, treated with more sodium tris-sodium succinate (0.310 g) and stirred in ice-water for 75 min. More sodium triethoxysulfonium borohydride (0.312 g) was added and the mixture was stirred for 70 minutes. More sodium triethoxysulfonate (0.619 g) was added and the mixture was stirred overnight to slowly slow down the cooling bath. The next day, more sodium triethoxysulfonium borohydride (0.307 g) was added and the mixture was stirred for 70 minutes. More acetic acid (0.055 mL) was added and the mixture was stirred for 80 minutes. The mixture was warmed to 40 ° C and stirred at this temperature for 25 minutes, after which it was concentrated in vacuo to a fast cerium oxide. The residue was purified by flash chromatography eluting EtOAc EtOAc The yield was 0.344 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ7.93 (s, 1H), 7.57-7.11 (m, 4H), 5.64-5.43 (m, 1H), 4.00-3.81 (m, 2H), 3.77-3.55 (m, 6H), 3.31 (seven peaks, J = 6.8 Hz, 1H), 3.13 - 2.88 (m, 2H), 2.57-2.35 (m, 4H), 2.16-1.94 (m, 2H), 1.74 -1.54 (m, 2H), 1.36 (d, J = 6.7 Hz, 6H), 0.84 (s, 9H), 0.01 (s, 6H).

f) (9-(3-(1-fluoro-2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -isopropylthiazol-4-yl)methanone

(9-(3-(2-(Terve-butyldimethylsilyloxy)-1-fluoroethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] a solution of undecyl-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 39, step e) (0.33 g) in THF (5 mL) Treatment (1 M in THF, 0.69 mL) was stirred at room temperature for 50 min. More tetrabutylammonium fluoride (1 M in THF, 0.69 mL) was added and the mixture was stirred for additional 80 min. The solution was then concentrated on a flash of EtOAc (EtOAc m.). The yield was 0.221 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.91 (d, J = 1.5 Hz, 1H), 7.36-7.19 (m, 4H), 5.45 (ddd, J = 48.5, 6.6, 3.7 Hz, 1H), 4.86-4.76 (m, 1H), 3.80-3.58 (m, 9H), 3.58-3.44 (m, 1H), 3.31 (seven peak, J = 6.8 Hz, 1H), 2.46-2.28 (m, 4H) ), 1.79-1.67 (m, 2H), 1.66-1.50 (m, 2H), 1.36 (d, J = 6.9 Hz, 6H).

g) 2-fluoro-2-(3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)methyl)phenyl)acetaldehyde

(9-(3-(1-fluoro-2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-iso A solution of propylthiazole-4-yl)methanone (Example 39, step f) (0.213 g) in EtOAc (EtOAc) . Dess-Martin periodinane (0.296 g) was added and the mixture was taken out from the cooling bath and stirred at room temperature for 20 minutes. More Dess-Martin periodinane (0.295 g) was added and the mixture was stirred at room temperature for an additional 30 minutes. The reaction was then quenched by the addition of saturated aqueous sodium thiosulfate (5 mL), saturated sodium bicarbonate (5 mL) and ethyl acetate (5 mL) and the mixture was stirred for 10 min. The mixture was then extracted twice with ethyl acetate and the combined organic extracts were washed with brine. Acetic acid (0.1 mL) was added, and the acidified extract was dried over anhydrous magnesium sulfate and concentrated in vacuo. The yield was 0.240 g.

m/z 460(M+H) ⁺ (APCI)

h) 7-((1R)-2-(2-fluoro-2-(3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)methyl)phenyl)ethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di- Trifluoroacetate

( R )-7-(3-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) A solution of 0.182 g) in MeOH (3 mL) was taken from acetic acid (0.039mL) and stirred for 5 min. Subsequent addition of 2-fluoro-2-(3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 a solution of -methyl)phenyl)acetaldehyde (Example 39, step g) (0.239 g) in MeOH (4 mL), and the mixture was stirred at room temperature for 5 min then cooled in ice-water. Treated with sodium triethoxysulfonate (0.146 g). The mixture was stirred on ice for 25 minutes, after which more sodium triacetoxyborohydride (0.444 g) was added. The mixture was then stirred over the weekend to allow it to slowly warm to room temperature. The mixture was concentrated in vacuo on the next Monday. The residue was dissolved in methanol (3mL) and water (1.5mL) of the mixture and by purified by preparative HPLC (Sunfire ^TM, Gradient: in 15 aqueous 0.2% TFA to 50% acetonitrile). The product containing fractions were concentrated in vacuo and co-evaporated three times from acetonitrile to give a colourless residue. The residue was triturated with EtOAc EtOAc (EtOAc:EtOAc)

m/z 670(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.94 (s, 1H), 7.60-7.45 (m, 4H), 6.93 (dd, J = 8.2, 2.3 Hz, 1H), 6.77 (d, J=7.9 Hz, 1H), 5.97 (dt, J=49.2, 9.7 Hz, 1H), 4.99-4.89 (m, 1H), 4.36-4.17 (m, 2H), 3.75-3.62 (m, 6H), 3.60 - 2.90 (m, 9H), 2.08-1.92 (m, 2H), 1.81-1.64 (m, 2H), 1.35 (d, J = 6.9 Hz, 6H). Six unobserved exchangeable protons.

Example 40 (R)-7-(2-(4-fluoro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-fluoro-5-(2-hydroxyethyl)benzaldehyde

Add 2,2,6,6-tetramethylpiperidine (3.0 g) dropwise to n-butyllithium (1.6 M in hexanes, 13 mL) in anhydrous tetrahydrofuran (12 mL) . After 10 min, 2-(4-fluorophenyl)ethanol (1.00 g) was added dropwise. The reaction mixture was stirred at -78 &lt;0&gt;C for 5 h then anhydrous DMF (3. The cooling bath was removed and the reaction mixture was allowed to warm to room temperature overnight. Ethyl acetate and aqueous HCl (2M) were added and the solution was separated. The ethyl acetate layer was washed with water, brine, dried over sodium sulfate, filtered and evaporated. Purification by silica gel chromatography, eluting with ethyl acetate: hexanes 1:1 eluted to give the subtitle compound. The yield was 0.43 g. ¹ H NMR (300MHz, CDCl ₃ ) δ 10.35 (s, 1H), 7.73 (dd, J = 6.4 & 2.0 Hz, 1H), 7.52-7.46 (m, 1H), 7.12 (dd, J = 10.0 & 8.4 Hz, 1H), 3.88 (t, J = 6.5 Hz, 2H), 2.90 (t, J = 6.5 Hz, 2H). An unobserved exchangeable proton.

b) (9-(2-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -isopropylthiazol-4-yl)methanone

Sodium triethoxysilane borohydride (0.339 g) was added to 2-fluoro-5-(2-hydroxyethyl)benzaldehyde (Example 40, step a) (0.135 g), (2-isopropylthiazole) 4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (0.339 g) and A stirred solution of acetic acid (0.046 mL) in NMP (7 mL). After 16 h, more 2-fluoro-5-(2-hydroxyethyl)benzaldehyde (Example 40, step a) (0.135 g) was added followed by sodium triethyloxyborohydride (0.339 g). After 2 h, the reaction mixture was diluted with EtOAc EtOAc m. The aqueous layer was extracted with ethyl acetate. The combined ethyl acetate solution was evaporated in vacuo. Purification by silica gel chromatography, eluting with ethyl acetate: triethylamine 10:1 to give the subtitle compound compound containing NMP. The sub-title compound was collected by applying a gum to a 10 g SCX cartridge eluting with methanol followed by 20% '880' aqueous ammonia in methanol. The solution was evaporated to dryness and the resulting gum was applied to a silica gel column eluting with ethyl acetate: triethylamine 10:1 to give a sub-title compound. The yield was 0.31 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.90 (s, 1H), 7.19 (d, J = 7.3 Hz, 1H), 7.13 - 7.07 (m, 1H), 6.97 (t, J = 9.3 Hz, 1H), 4.28-4.24 (m, 1H), 3.71-3.55 (m, 6H), 3.46 (s, 2H), 3.35-3.26 (m, 1H), 3.00 (s, 2H), 2.70 (t , J = 6.7 Hz, 2H), 2.45-2.26 (m, 4H), 1.74-1.65 (m, 2H), 1.59-1.49 (m, 2H), 1.36 (d, J = 7.0 Hz, 6H).

c) 2-(4-Fluoro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)methyl)phenyl)acetaldehyde

Add Dess-Martin periodinane (0.284 g) to (9-(2-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5 (undecyl-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 40, step b) (0.21 g) and trifluoroacetic acid (0.046 mL) in DCM (5 mL) Stir in the solution. After 1 h, ethyl acetate (30 mL) was added followed by a mixture of saturated sodium thiosulfate (5 mL) and saturated sodium bicarbonate (5 mL). The reaction mixture was thoroughly shaken and separated. The ethyl acetate solution was washed with saturated sodium bicarbonate solution, water and brine. Acetic acid (0.07 mL) was added, the solution was dried over sodium sulfate, filtered and evaporated in vacuo. The yield was 0.2 g. Use directly.

m/z 460(M+H) ⁺ (APCI)

d) (R)-7-(2-(4-fluoro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

Acetic acid (0.037 mL) was added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.17 g) and 2-(4-fluoro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diaza A stirred solution of spiro[5.5]undecane-9-yl)methyl)phenyl)acetaldehyde (Example 40, step c) (0.20 g) in methanol (8 mL). After 1 min, sodium cyanoborohydride (0.10 g) was added. After 3 h, the mixture was evaporated w~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ The aqueous layer was extracted with ethyl acetate. The THF and ethyl acetate solutions were combined and evaporated in vacuo. The gum was dissolved in methanol and purified by preparative HPLC (Sunfire ^(TM) , gradient: 10 to 35% acetonitrile in 0.2% aqueous TFA). The fractions containing the pure product were combined and evaporated to dryness. The title compound was obtained as a white solid. The yield was 0.126 g.

m/z 670(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ 11.33-11.22 (m, 1H), 7.94 (s, 1H), 7.46-7.36 (m, 2H), 7.25 (t, J = 9.0Hz, 1H ), 6.93 (d, J = 8.4 Hz, 1H), 6.77 (d, J = 8.4 Hz, 1H), 4.93-4.88 (m, 1H), 4.30-4.20 (m, 2H), 3.70 (s, 4H) , 3.66 (s, 2H), 3.35-2.93 (m, 11H), 2.05-1.95 (m, 2H), 1.83-1.67 (m, 2H), 1.35 (d, J = 7.0 Hz, 6H). Five unobserved exchangeable protons.

Example 41 (R)-7-(2-(3-fluoro-5-((4-(2-phenylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] eleven Cycloalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(3-(bromomethyl)-5-fluorophenyl)acetic acid

Benzoyl peroxide (0.5 g) was added to a stirred mixture of 2-(3-fluoro-5-methylphenyl)acetic acid (5.19 g) and NBS (6.04 g) in dichloromethane (100 mL). The resulting mixture was heated under reflux for 5 hours. The reaction mixture was cooled to room temperature, then washed twice with water, dried over sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on silica gel using 1% acetic acid and 17% ethyl acetate in isohexane as solvent. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 4.3 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 7.22 - 7.17 (m, 2H), 7.09 - 7.05 (m, 1H), 4.68 (s, 2H), 3.61 (s, 2H). An unobserved exchangeable proton.

b) 2-(3-(bromomethyl)-5-fluorophenyl)ethanol

Borane-methylthio complex (2M in THF, 17.4 mL) was added dropwise to 2-(3-(bromomethyl)-5-fluorophenyl)acetic acid over 10 min (Example 41, step a (4.3 g) in THF (60 mL) in EtOAc. The mixture was stirred at 0 ° C for 10 minutes and then at 20 ° C for 1 hour. The reaction mixture was added dropwise with methanol to stop the reaction and the solvent was removed under reduced pressure. The crude product was purified by flash chromatography on silica gel eluting with 30% ethyl acetate in isohexane. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 3.7 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ7.15 (s, 1H), 7.14-7.09 (m, 1H), 7.05-6.99 (m, 1H), 4.66 (s, 2H), 3.61 (t, J = 6.8 Hz, 2H), 2.72 (t, J = 6.7 Hz, 2H). An unobserved exchangeable proton.

c) 2,2,2-trifluoro-1-(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] Undecyl-4-yl)ethanone

2-(3-(Bromomethyl)-5-fluorophenyl)ethanol (Example 41, step b) (3.6 g) and 2,2,2-trifluoro-1-(1-oxa-4, A solution of 9-diazaspiro[5.5]undecane-4-yl)ethanone trifluoroacetate (Example 12, step d) (5.66 g) in EtOAc (EtOAc) 5.38 mL) and the mixture was stirred at 20 ° C for 20 hours. The solvent was evaporated under reduced pressure and the residue was partitioned between ethyl acetate and brine. The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 4.9 g.

m/z 405(M+H) ⁺ (APCI)

d) (9-(3-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -phenylthiazole-4-yl)methanone

2,2,2-trifluoro-1-(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] A solution of the carboxane-4-yl)ethanone (Example 41, step c) (0.21 g) in MeOH (3 mL)EtOAc. The mixture was evaporated to dryness under reduced pressure. The residue was dissolved in DMF (7 mL) EtOAc (EtOAc (EtOAc:EtOAc) Stir for 1 hour. The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The yield was 0.27 g.

m/z 496 (M+H) ^ten (APCI)

e) (R)-7-(2-(3-Fluoro-5-((4-(2-phenylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

(9-(3-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-benzene A solution of the thiothiazol-4-yl)methanone (Example 41, step d) (0.25 g) in dichloromethane (20 mL) eluting with trifluoroacetic acid (0.039 mL) (0.278 g) was treated, and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.029 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.199 g) and acetic acid (0.029 mL) in MeOH (15 mL). The mixture was cooled in an ice-bath and treated with sodium cyanoborohydride (0.063 g). The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of brine (10 mL) and saturated sodium bicarbonate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.19 g.

m/z 704 (M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D _{6 -} DMSO, 90 ° C) δ 11.28 (s, 1H), 8.14 (s, 1H), 7.96-7.91 (m, 2H) ), 7.54-7.49 (m, 3H), 7.28-7.18 (m, 3H), 6.93 (d, J = 8.5 Hz, 1H), 6.77 (d, J = 8.5 Hz, 1H), 4.94 - 4.87 (m, 1H), 4.28 (s, 2H), 3.83-3.66 (m, 6H), 3.26 (t, J = 7.9 Hz, 2H), 3.21-2.99 (m, 8H), 2.10-2.00 (m, 2H), 1.84 -1.69 (m, 2H). Five unobserved exchangeable protons.

Example 42 (R)-7-(2-(3-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzene well [d]thiazole-2(3H)-one di-trifluoroacetate

a) (9-(3-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -isopropylthiazol-4-yl)methanone

2,2,2-trifluoro-1-(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] A solution of the carboxane-4-yl)ethanone (Example 41, step d) (0.21 g) in MeOH (3 mL)EtOAc. The mixture was evaporated to dryness under reduced pressure. The residue was dissolved in DMF (7 mL) EtOAc (EtOAc (EtOAc) Stir at °C for 1 hour. The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The yield is 0.25 g.

m/z 462(M+H) ⁺ (APCI)

b) (R)-7-(2-(3-Fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

(9-(3-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-iso A solution of propylthiazole-4-yl)methanone (Example 42, step a) (0.23 g) in DCM (20 mL 0.275 g) was treated, and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.029 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.196 g) and acetic acid (0.029 mL) in methanol (15 mL). The mixture was cooled in an ice-bath and treated with sodium cyanoborohydride (0.063 g). The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.15 g.

m/z670(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.27 (s, 1H), 7.94 (s, 1H), 7.28-7.16 (m, 3H), 6.93 (d, J = 8.5 Hz, 1H) , 6.77 (d, J = 8.2 Hz, 1H), 4.94 - 4.88 (m, 1H), 4.27 (s, 2H), 3.70 (s, 4H), 3.66 (s, 2H), 3.32-3.23 (m, 3H) ), 3.17-2.99 (m, 8H), 2.07-1.96 (m, 2H), 1.81-1.69 (m, 2H), 1.35 (d, 6H). Five unobserved exchangeable protons.

Example 43 (R)-7-(2-(2-(5-((4-(2-T-butylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Undecane-9-yl)methyl)thiophen-3-yl)ethylamino)-1-hydroxyethyl)-4-hydroxybenzene[d]thiazole-2(3H)-one di-three Fluoroacetate

a) 1-(9-((4-(2-(Tern-butyldimethyl decyloxy)ethyl)thiophen-2-yl)methyl)-1-oxa-4,9- Diazaspiro[5.5]undecane-4-yl)-2,2,2-trifluoroethyl ketone

2,2,2-Trifluoro-1-(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)ethanone trifluoroacetate (Example 12, step d ) (1.084 g) was added to 4- (2- (third - butyldimethylsilyl silicon alkyloxy) ethyl) thiazole-2-carbaldehyde and thiophene 3- (2- (third - butyldimethyl 4:1 mixture of decyloxy)ethyl)thiophene-2-carboxaldehyde (Example 27, step b) (1.0 g) and AcOH (0.16 mL) in N-methyl-2-pyrrolidone (15 mL) In the stirred solution. After 5 min, sodium triethoxysulfonium borohydride (1.57 g) was added. After 16 h, water was added and the mixture was extracted with ethyl acetate. The ethyl acetate layer was washed three times with water and evaporated in vacuo. Purification by silica gel chromatography, eluting with ethyl acetate: isohexane:triethylamine 1:20:1. The yield was 1.04 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ6.87 (s, 1H), 6.75 (d, J = 3.7Hz, 1H), 3.79-3.72 (m, 4H), 3.65 (s, 3H), 3.59-3.55 ( m, 1H), 3.52 (s, 1H), 3.37 (s, 1H), 2.76 (t, J = 6.9 Hz, 2H), 2.68-2.61 (m, 1H), 2.57-2.50 (m, 1H), 2.46 -2.38 (m, 1H), 2.31 (t, J = 11.5 Hz, 1H), 1.89-1.75 (m, 2H), 1.68-1.52 (m, 2H), 0.87 (s, 9H), 0.00 (s, 6H) ).

b) 9-((4-(2-(Third-butyldimethylsilyloxy)ethyl)thiophen-2-yl)methyl)-1-oxa-4,9-diaza Snail [5.5] undecane

The '880' aqueous ammonia solution (1.5mL) was added to 1- (9 - ((4- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethyl) thiophen-2-yl) methyl) -1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)-2,2,2-trifluoroethyl ketone (Example 43, step a) (1.0 g) Stirred in methanol (5 mL). After 16 h, the reaction mixture was evaporated to dryness. Acetonitrile was added and the solution was evaporated to dryness in vacuo. The yield was 0.78 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ6.94 (s, 1H), 6.87 (s, 1H), 3.91-3.85 (m, 2H), 3.78 (t, J = 7.1Hz, 2H), 3.72-3.68 ( m, 2H), 2.93-2.82 (m, 4H), 2.80-2.73 (m, 4H), 2.62-2.53 (m, 2H), 2.11-2.01 (m, 2H), 1.76-1.64 (m, 2H), 0.87 (s, 9H), 0.00 (s, 6H) + 1 unobserved exchangeable protons.

c) (9-((4-(2-(Third-butyldimethylsilyloxy)ethyl)thiophen-2-yl)methyl)-1-oxa-4,9-diaza Heterospiro[5.5]undecane-4-yl)(2-tris-butylthiazol-4-yl)methanone

The HATU (0.306g) was added to 2- tertiary - butyl-4-carboxylic acid (0.16g), 9 - (( 4- (2- ( tertiary - butyl dimethyl silicone alkyloxy) ethyl Thiophen-2-yl)methyl)-1-oxa-4,9-diazaspiro[5.5]undecane (Example 43, step b) (0.300 g) and triethylamine (0.41 mL) ) in a stirred solution in DMF (3 mL). After 1 h, the reaction mixture was partitioned between water and ethyl acetate. The ethyl acetate layer was washed twice with water and brine, dried over magnesium sulfate, filtered and evaporated. Purification by silica gel chromatography, eluting with ethyl acetate: hexanes: triethylamine 12: 90:10 to give the subtitle compound. The yield was 0.3 g.

_{^{1 H NMR (300MHz, CDCl 3}} ) δ7.82 (s, 1H), 6.85 (s, 1H), 6.74 (s, 1H), 4.01-3.92 (m, 1H), 3.92-3.84 (m, 1H), 3.82-3.71 (m, 7H), 3.69-3.58 (m, 3H), 2.79-2.71 (m, 2H), 2.60-2.28 (m, 4H), 1.93-1.79 (m, 2H), 1.44 (s, 9H) ), 0.87 (s, 9H), -0.01 (s, 6H).

d) (2-t-butylthiazol-4-yl)(9-((4-(2-hydroxyethyl))thiophen-2-yl)methyl)-1-oxa-4,9-di Azaspiro[5.5]undecane-4-yl)methanone

The TBAF (1.5mL of a 1M solution in THF) was added to (9 - ((4- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethyl) thiophen-2-yl) methyl) oxa-4,9-diaza-spiro [5.5] undecane-4-yl) (2-tertiary - butyl-thiazol-4-yl) methanone (example 43, step c) (0.300 g) in a stirred solution of THF (3 mL). After 1 h, the reaction evaporated to a gum. Purification by silica gel chromatography, eluting with ethyl acetate: triethylamine 20:1 affords the subtitle compound. The yield was 0.22 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 8.00 (s, 1H), 7.01 (s, 1H), 6.84 - 6.73 (m, 1H), 4.59 (t, J = 5.3 Hz, 1H), 3.77- 3.48 (m, 10H), 2.64 (t, J = 7.0 Hz, 2H), 2.54-2.16 (m, 4H), 1.74-1.63 (m, 2H), 1.58-1.43 (m, 2H), 1.41 (s, 9H).

e) 2-(5-((4-(2-T-butylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl) )methyl)thiophen-3-yl)acetaldehyde

Add Dess-Martin periodine (0.316 g) to (2-t-butylthiazol-4-yl)(9-((4-(2-hydroxyethyl))thiophen-2-yl)methyl --1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone (Example 43, step d) (0.23 g) and TFA (0.05 mL) in DCM ( In a stirred solution in 5 mL). After 40 min, the reaction mixture was crystallisjjjjjjjjjjjjjjjjjjj The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. The yield was 0.23 g. Use directly.

m / z 462 (M + H ) + (APCI)

f) (R)-7-(2-(2-(5-((4-(2-T-butylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)thiophen-3-yl)ethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one -trifluoroacetate

Acetic acid (0.039 mL) was added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, ((4- (2-third - - Example 1, step d) (0.180g) and 2- (5-butyl-thiazole-4- carbonyl embodiment yl) -l-oxa-4,9-diazaspiro [ 5.5] a stirred solution of undecane-9-yl)methyl)thiophen-3-yl)acetaldehyde (Example 43, step e) (0.230 g) in MeOH (8 mL). After 1 min, sodium cyanoborohydride (0.125 g) was added. After 3h, the reaction mixture was filtered and purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in acetonitrile 10 to 35%) by. The fractions containing the pure product were combined and evaporated to dryness. The title compound was obtained as a white solid. The yield was 0.14 g.

m/z 672(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ11.67 (s, 1H), 10.24 (s, 1H), 8.91-8.71 (m, 2H), 8.04 (s, 1H), 7.46 (s, 1H), 7.18(s,1H), 6.93(d,J=8.4Hz,1H),6.77(d,J=8.4Hz,1H),6.54-6.44(m,1H),4.93-4.86(m,1H),4.68 -4.51 (m, 2H), 3.82-3.47 (m, 8H), 3.32-3.15 (m, 4H), 3.12-2.87 (m, 5H), 2.15-2.07 (m, 2H), 1.83-1.54 (m, 2H), 1.39 (s, 9H).

Example 44 (R)-5-(2-(3-((2,2-difluoro-4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetate

a) 2,2-difluoro-4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid third Butyl ester

Add HATU (1.184 g) to 2,2-difluoro-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester (Example 37, step c (0.7 g) and 2-isopropylthiazole-4-carboxylic acid (0.41 g) and triethylamine (1 mL) in DMF (12 mL) in 20 ° C, and the mixture was stirred for 1 hour. The mixture was partitioned between EtOAc and EtOAc. The crude product was purified by flash chromatography on silica gel eluting with 30% ethyl acetate in isohexane. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.78 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.11 (s, 1H), 4.29 - 4.20 (m, 2H), 4.03-3.98 (m, 2H), 3.58-3.51 (m, 2H), 3.37-3.30 (m, 1H), 3.26-3.17 (m, 2H), 1.81-1.74 (m, 2H), 1.66-1.57 (m, 2H), 1.40 (s, 9H), 1.35 (d, 6H)

b) (2,2-difluoro-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-isopropylthiazol-4-yl)methanone Fluoroacetate

2,2-Difluoro-4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester (Example 44, step a) (0.78 g) <RTI ID=0.0></RTI> Toluene (40 mL) was added and the solvent was evaporated under reduced pressure. The residue was azeotroped twice with acetonitrile to give the subtitle compound. The yield was 0.8 g.

m/z 346(M+H) ⁺ (APCI)

_{^{1 H NMR (300MHz, D 6}} -DMSO, 90 ℃) δ8.57 (s, 2H), 8.16 (s, 1H), 4.39-4.22 (m, 2H), 4.04 (s, 2H), 3.39-3.28 ( m, 1H), 3.27-3.19 (m, 2H), 3.16-3.04 (m, 2H), 2.07-1.97 (m, 2H), 1.93-1.80 (m, 2H), 1.37 (d, 6H).

c) (2,2-difluoro-9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl (2-isopropylthiazol-4-yl)methanone

2-(3-(Bromomethyl)phenyl)ethanol (Example 6, step a) (0.21 g) was added to (2,2-difluoro-1-oxa-4,9-diazaspiro) [5.5] undecane-4-yl)(2-isopropylthiazol-4-yl)methanone trifluoroacetate (Example 44, step b) (0.40 g) and triethylamine (0.37 mL) In a mixture of acetonitrile (15 mL). The reaction mixture was stirred at 20 ° C for 2 hours. The solvent was evaporated under reduced pressure and EtOAc evaporated m. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.40 g.

m/z 480(M+H) ⁺ (APCI)

d) (R)-5-(1-(Third-butyldimethylmethylalkyloxy)-2-(3-((2,2-difluoro-4-(2-isopropylthiazole)- 4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)-8-hydroxyquinoline-2 (1H)-ketone

(2,2-Difluoro-9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)( a solution of 2-isopropylthiazol-4-yl)methanone (Example 44, step c) (0.38 g) in DCM (20 mL) Treated with iodine (0.437 g) and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.045 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3mL) and added to (R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8-hydroxy-quinolin-2 (1H)-ketone (WO 2004106333) (0.345 g) in MeOH (15 mL). The mixture was cooled to 0 ° C and sodium triethoxysulfonate (0.252 g) was added in one portion. The reaction mixture was stirred at 20 ° C for 3 hours. The solvent was removed under reduced pressure and the residue was crystalljjjjjjjjjjjjjjj The crude product was purified by flash chromatography on silica gel using 10% methanol in dichloromethane containing 1% &quot;880" The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.32 g.

m/z796(M+H) ⁺ (APCI)

e) (R)-5-(2-(3-((2,2-difluoro-4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetate

Triethylamine trihydrofluoride (0.082 mL) in methanol (2 mL) was added to ( R )-5-(1-(t-butyldimethylmethylalkyloxy)-2-(3) -((2,2-difluoro-4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl) Methyl)phenethylamino)ethyl)-8-hydroxyquinolin-2(1H)-one (Example 44, step d) (0.32 g) in THF (8 mL) The mixture was allowed to stand at 20 ° C for 18 hours. The solvent was removed under reduced pressure and the crude material was purified by preparative HPLC (Sunfire ^(s) , gradient: 10 to 40% acetonitrile in 0.2% aqueous TFA). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.21 g.

m/z682(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.18-8.11 (m, 2H), 7.44 - 7.31 (m, 4H), 7.14 (d, J = 8.2 Hz, 1H), 6.99 (d, J=7.9 Hz, 1H), 6.55 (d, J = 10.0 Hz, 1H), 5.37-5.31 (m, 1H), 4.31 (s, 2H), 4.23 (s, 2H), 4.01 (s, 2H), 3.40-2.98 (m, 11H), 2.08-1.99 (m, 2H), 1.98-1.88 (m, 2H), 1.36 (d, 6H). Six unobserved exchangeable protons.

Example 45 (R)-7-(2-(2,6-difluoro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(2,6-difluorophenyl)ethanol

The borane dimethyl sulfide solution complex (2M in THF, 26 mL) was carefully added to a solution of 2-(2,6-difluorophenyl)acetic acid (3 g) in THF (50 mL) at 0 ° C in. The reaction was then allowed to warm to RT and stirred for 3 h. The reaction was cooled in an ice bath and the reaction was quenched with methanol (10 mL). The solvent was evaporated and the residue was purified EtOAc EtOAc elut elut The fractions containing the product are combined and evaporated to give the clear oil of the sub-title compound. The yield was 2.2 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.20-7.13 (m, 1H), 6.94-6.82 (m, 2H), 3.85 (t, J = 6.8Hz, 2H), 2.97 (t, J = 6.7Hz, 2H). An unobserved exchangeable proton.

b) 2,4-difluoro-3-(2-hydroxyethyl)benzaldehyde

2,2,6,6-Tetramethylpiperidine (5 mL) was added to a solution of butyl lithium (1.6M in hexanes, 19 mL) in THF (25 mL). A solution of 2-(2,6-difluorophenyl)ethanol (Example 45, Step a) (1.6 g) in THF (25 mL). Then DMF (3.9 mL) was added and the mixture was stirred at -70 ° C for 1 h. The mixture was then allowed to warm to RT and stirred for 70 h. The reaction was quenched with EtOAc (2M, 50 mL)EtOAc. The aqueous phase was extracted with ethyl acetate (2×50 mL). The combined organics were washed with brine (100 mL) dried over magnesium sulfate. The residue was purified by silica gel chromatography eluting with 4:1 to 2:1 isohexane: diethyl ether gradient. The fractions containing the product are combined and evaporated. The oil was dissolved in EtOAc (EtOAc) (EtOAc) The yield is 1.1g.

_{^{1 H NMR (300MHz, CDC1 3}} ) δ10.28 (s, 1H), 7.84-7.74 (m, 1H), 7.05-6.95 (m, 1H), 3.89 (t, J = 6.6Hz, 2H), 3.01 ( t, J = 6.6 Hz, 2H). An unobserved exchangeable proton.

c) (9-(2,4-Difluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl (2-isopropylthiazol-4-yl)methanone

Add 2,4-difluoro-3-(2-hydroxyethyl)benzaldehyde (Example 45, step b) (0.35 g) to (2-isopropylthiazol-4-yl)(1-oxa -4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (0.67 g) and acetic acid (0.09 mL) in N-methyl In a solution of 2-pyrrolidone (1 mL). The resulting mixture was stirred for 1 h then quenched by EtOAc EtOAc (EtOAc &lt The resulting mixture was stirred overnight, diluted with EtOAc (20 mL) and applied to EtOAc (EtOAc &lt The column was washed with acetonitrile (50 mL) and dissolved in 10% &lt;RTI ID=0.0&gt;&gt; Evaporation of the eluent, azeotrope with toluene and purification by gel chromatography, eluting with 77.5:17.5:5 isohexane:ethyl acetate:triethylamine to 95:5 ethyl acetate:triethylamine Yellow gum. The yield was 0.74 g.

m/z 480(M+H) ⁺ (APCI)

¹ H NMR (300 MHz, D _{6 -} DMSO) δ 8.00 (s, 1 H), 7.32 - 7.16 (m, 1H), 7.00 (t, J = 8.7 Hz, 1H), 4.80 (t, J = 5.5 Hz, 1H), 3.78-3.38 (m, 11H), 2.77 (t, J = 7.0 Hz, 2H), 2.46-2.26 (m, 4H), 1.76-1.40 (m, 4H), 1.35 (d, J = 6.9 Hz) , 6H).

d) 2-(2,6-Difluoro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undeccarbon Alkan-9-yl)methyl)phenyl)acetaldehyde

Add TFA (0.11 mL) to (9-(2,4-difluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] eleven Carboxy-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 45, step c) (0.7 g) in DCM (5 mL) in EtOAc Stir for 5 min. Dess-Martin periodinane (0.93 g) was then added and the mixture was stirred at RT for 45 min. Saturated sodium thiosulfate solution (5 mL), saturated sodium bicarbonate (5 mL) and ethyl acetate (20 mL) The aqueous layer was separated and extracted with ethyl acetate (20 mL). The combined organics were washed with brine, EtOAc (EtOAc)EtOAc. The yield was 0.64 g.

m/z 478(M+H) ⁺ (APCI)

e) (R)-7-(2-(2,6-difluoro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoro Acetate

2-(2,6-Difluoro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane) A solution of 9-yl)methyl)phenyl)acetaldehyde (Example 45, step d) (0.323 g) in methanol (3 mL) was added to (R)-7-(2-amino-1-hydroxyl Ethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.23 g) and acetic acid (0.034 mL) in methanol (2 mL) In the mixture. The resulting mixture was stirred for 5 min and then cooled to 0 °C. Sodium cyanoborohydride (0.057 g) was added and the mixture was warmed to RT and stirred for 2 h. The solvent was evaporated and the residue was purified by chromatography eluting eluting eluting eluting elution The combined product-containing fractions were dissolved, evaporated and purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile) to borrow. The title compound was obtained as a white solid. The yield was 0.21 g.

m/z 688(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.27 (s, 1H), 7.94 (s, 1H), 7.62-7.53 (m, 1H), 7.23-7.14 (m, 1H), 6.93 ( d, J = 8.2 Hz, 1H), 6.78 (d, J = 8.2 Hz, 1H), 4.92 (dd, J = 8.1, 4.7 Hz, 1H), 4.31 (s, 2H), 3.75 - 3.61 (m, 6H) ), 3.34 - 3.02 (m, 11H), 2.10 - 1.95 (m, 2H), 1.86-1.71 (m, 2H), 1.34 (d, J = 6.7 Hz, 6H). Five unobserved exchangeable protons.

Example 46

(R)-5-(2-(2,6-difluoro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetate

2-(2,6-Difluoro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane) A solution of 9-yl)methyl)phenyl)acetaldehyde (Example 45, step d) (0.29 g) in methanol (3 mL) was added to ( R )-5-(2-amino-1-( third - butyldimethyl silicon alkyloxy) ethyl) -8-hydroxy-quinolin -2 (1H) - one (WO 2004106333) (0.20g) and acetic acid (0.034 mL) in methanol (3mL) in the In the mixture. The resulting mixture was stirred for 5 min and then cooled to 0 °C. Sodium cyanoborohydride (0.057 g) was then added and the mixture was warmed to RT and stirred for 2 h. The solvent was evaporated and the residue was purified by EtOAc EtOAc (EtOAc:EtOAc:EtOAc The fractions containing the product are combined and evaporated. The residue was dissolved in THF (3 mL) EtOAc. The reaction was concentrated and the residue was purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile) to borrow. The title compound was obtained as a white solid. The yield was 0.23 g.

m/z 682(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.19 (d, J = 10.0 Hz, 1H), 7.94 (s, 1H), 7.65-7.51 (m, 1H), 7.24-7.10 (m, 2H) ), 7.01 (d, J = 7.9 Hz, 1H), 6.53 (d, J = 10.0 Hz, 1H), 5.37 (dd, J = 8.6, 4.0 Hz, 1H), 4.34 (s, 2H), 3.75-3.59 (m, 6H), 3.34 - 3.03 (m, 11H), 2.11 - 1.95 (m, 2H), 1.88 - 1.72 (m, 2H), 1.34 (d, J = 6.9 Hz, 6H). Six unobserved exchangeable protons.

Example 47

(R)-7-(2-(2-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(5-(bromomethyl)-2-fluorophenyl)ethanol

Dibenzoyl peroxide (0.14 g) was added to a solution of NBS (1.53 g) and 2-(2-fluoro-5-methylphenyl)acetic acid (1.45 g) in DCM (50 mL). The mixture was heated under reflux for 12 h. The solvent was evaporated and the white solid was dissolved in ethyl acetate (100 mL) The layers were separated and EtOAc (EtOAc)EtOAc The resulting white solid was redissolved in tetrahydrofuran (25 mL) and cooled in ice. The borane dimethyl sulfide solution complex (2M in THF, 13 mL) was carefully added and the mixture was then warmed to RT and stirred for 2 h. The reaction was cooled in an ice bath and the reaction was carefully quenched with methanol. Once the bubbling was stopped, the solvent was evaporated and the residue was purified eluting with EtOAc EtOAc EtOAc EtOAc The yield was 1.35 g.

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 7.32-7.21 (m, 2H), 7.04-6.97 (m, 1H), 4.46 (s, 2H), 3.87 (t, J = 6.5Hz, 2H), 2.93-2.87 (m, 2H). An unobserved exchangeable proton.

b) (9-(4-Fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -isopropylthiazol-4-yl)methanone

2-(5-(Bromomethyl)-2-fluorophenyl)ethanol (Example 47, step a) (0.16 g) was added to (2-isopropylthiazol-4-yl)(1-oxa) -4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (0.3 g) and triethylamine (0.3 mL) in acetonitrile ( In a solution of 10 mL). The resulting mixture was stirred overnight, diluted with EtOAc (20 mL) and applied to EtOAc (EtOAc) The column was washed with acetonitrile (50 mL) and dissolved in 10% &lt;RTI ID=0.0&gt;&gt; The eluent was evaporated, azeotroped with toluene and purified by silica gel chromatography eluting with 77.5.17.5:5 isohexane:ethyl acetate:triethylamine to 95:5 ethyl acetate:triethylamine gradient. The fractions containing the product are combined and evaporated to give the sub-title compound as a yellow gum. The yield was 0.22 g.

m/z 462(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.91 (s, 1H), 7.22 - 7.14 (m, 1H), 7.14 - 7.06 (m, 1H), 7.00 (dd, J = 10.0, 8.3 Hz, 1H), 4.40 (t, J = 5.3 Hz, 1H), 3.72-3.52 (m, 8H), 3.43 - 3.23 (m, 3H), 2.74 (t, J = 7.0 Hz, 2H), 2.42 - 2.23 (m, 4H), 1.75-1.65 (m, 2H), 1.59-1.47 (m, 2H), 1.36 (d, J = 6.9 Hz, 6H).

c) (R)-7-(2-(2-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl 4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

Adding trifluoroacetic acid (0.033 mL) to (9-(4-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undec carbon Alkyl-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 47, step b) (0.2 g) in DCM (5 mL) The mixture was stirred for 5 min, then Dess-Martin periodinane (0.28 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (1 mL), sat. sodium bicarbonate (1 mL) and ethyl acetate (5 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (5 mL). The combined organic solution was washed with brine (5 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (2 mL) followed by acetic acid (0.025 mL) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.17 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.04 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 30% acetonitrile). The title compound was obtained as a white solid. The yield was 0.14 g.

m/z 670(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.27 (s, 1H), 7.94 (s, 1H), 7.52-7.42 (m, 2H), 7.25 (t, J = 9.2Hz, 1H) , 6.94 (d, J = 8.2 Hz, 1H), 6.78 (d, J = 8.2 Hz, 1H), 4.93 (dd, J = 7.9, 4.6 Hz, 1H), 4.29 (s, 2H), 3.80-3.58 ( m, 6H), 3.34 - 2.95 (m, 11H), 2.11-1.95 (m, 2H), 1.87-1.64 (m, 2H), 1.34 (d, J = 6.7 Hz, 6H). Five unobserved exchangeable protons.

Example 47A

(R)-7-(2-(2-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one a)2-(5-(bromo) Methyl)-2-fluorophenyl)ethanol

Dibenzoyl peroxide (1 g) was added to a solution of NBS (10.6 g) and 2-(2-fluoro-5-methylphenyl)acetic acid (10 g) in DCM (250 mL). Heated under reflux for 12 h. The solvent was evaporated and the white solid was dissolved in ethyl acetate (250mL) The layers were separated and the organic phase washed with EtOAc EtOAc EtOAc EtOAc The resulting white solid was redissolved in tetrahydrofuran (150 mL) and cooled in ice. The borane dimethyl sulfide solution complex (2M in THF, 89 mL) was carefully added and the mixture was then warmed to RT and stirred overnight. The reaction was cooled in an ice bath and the reaction was carefully quenched with methanol. Once bubbling ceased, the solvent was evaporated and the residue was triturated with a 4:1 mixture of isohexane and diethyl ether. Purification by silica gel chromatography, eluting with 9:1 to 4:1 ethyl acetate: EtOAc (EtOAc) The yield was 6.5 g. _{^{1 H NMR (300MHz, CDCl 3}} ) δ 7.32-7.21 (m, 2H), 7.04-6.97 (m, 1H), 4.46 (s, 2H), 3.87 (t, J = 6.5Hz, 2H), 2.93-2.87 (m, 2H). An unobserved exchangeable proton.

b) (9-(4-Fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -isopropylthiazol-4-yl)methanone

2-(5-(Bromomethyl)-2-fluorophenyl)ethanol (Example 47A, Step a) (5.2 g) was added to (2-isopropylthiazol-4-yl)(1-oxa -4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (9.4 g) and potassium carbonate (6.8 g) in ethanol (75 mL) In the suspension in). The resulting mixture was stirred overnight and filtered. The residue was washed with ethanol (50 mL) and the combined filtrate and washings were evaporated. The residue was partitioned between EtOAc (EtOAc)EtOAc. Purified by silica gel chromatography, eluting with 95:5 ethyl acetate:triethylamine gradient. The fractions containing the product are combined and evaporated to give the sub-title compound as a yellow gum. The yield was 7.9 g.

m/z 462 (M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D _{6 -} DMSO, 90 ° C) δ 7.91 (s, 1H), 7.22 - 7.14 (m, 1H), 7.14 - 7.06 (m) , 1H), 7.00 (dd, J = 10.0, 8.3 Hz, 1H), 4.40 (t, J = 5.3 Hz, 1H), 3.72-3.52 (m, 8H), 3.43 - 3.23 (m, 3H), 2.74 ( t, J = 7.0 Hz, 2H), 2.42 - 2.23 (m, 4H), 1.75-1.65 (m, 2H), 1.59-1.47 (m, 2H), 1.36 (d, J = 6.9 Hz, 6H).

c) (R)-7-(2-(2-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one

Adding trifluoroacetic acid (1.32 mL) to (9-(4-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undec carbon Alkyl-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 47A, step b) (7.9 g) in DCM (200 mL) The mixture was stirred for 5 min, then Dess-Martin periodinane (12.3 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (100 mL), saturated sodium bicarbonate solution (100 mL) and ethyl acetate (500 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (100 mL). The combined organic solution was washed with EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (140mL), followed by addition of (R) -7- (2- amino-1-hydroxyethyl) -4-hydroxybenzo [d] thiazol -2 (3H) - one hydrochloride ( WO 2007027134, Example 1, step d) (4.5 g) and the mixture was stirred for 5 min before cooling in an ice bath. Sodium cyanoborohydride (1.6 g) was then added and the mixture was warmed to RT and stirred overnight. The reaction mixture was concentrated in EtOAc (EtOAc)EtOAc. The layers were separated and dried with sodium sulfate, filtered, evaporated and evaporated Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: "880" aqueous ammonia. The title compound was obtained as a white solid. Yield 4.1 gm/z 670 (M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.27 (s, 1H), 7.94 (s, 1H), 7.52-7.42 (m, 2H), 7.25 (t, J = 9.2 Hz, 1H), 6.94 (d, J = 8.2 Hz, 1H), 6.78 (d, J = 8.2 Hz, 1H), 4.93 (dd, J = 7.9, 4.6 Hz, 1H), 4.29 (s, 2H), 3.80-3.58 (m, 6H), 3.34 - 2.95 (m, 11H), 2.11-1.95 (m, 2H), 1.87-1.64 (m, 2H), 1.34 (d, J = 6.7 Hz, 6H). Five unobserved exchangeable protons.

real Example 47B

(R)-7-(2-(2-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di(1S)-(+)- 10-camphorsulfonate

Add 1S-(+)-camphorsulfonic acid (41 mg) to (R)-7-(2-(2-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-) Oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole- 2(3H)-one (Example 47A) (59 mg) in EtOAc (5 mL), and evaporated to dryness to give (R)-7-(2-(2-fluoro-5-) ((4-(2-Isopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl)phenethylamino An amorphous white solid of 1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one dicamphorsulfonate. The yield is 0.1 g.

Add isopropyl alcohol (1 mL) to (R)-7-(2-(2-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9) -diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one The dicamphorsulfonate (20 mg) was added and the resulting clear solution was stirred for 2 days. A white solid formed and the suspension was stirred for another 5 days. Centrifugal filtration of the solid and drying under high vacuum yields (R)-7-(2-(2-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa) -4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2 ( 3H)-White di-camphorsulfonate as a white crystalline solid. The yield is 5 mg.

(R)-7-(2-(2-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one (4.3 g, 6.42 mmol) (implementation Example 47A), a mixture of (1S)-(+)-10-camphorsulfonic acid (2.98 g, 12.84 mmol) and iso -propanol (300 mL) was heated at 50 ° C until a clear solution formed, and seed crystals were added to make Cool to RT and stir for 4 days. Solid was isolated by filtration to iso - propanol (100 mL), diethyl ether (2 × 200mL) and washed suction dried to give the title compound as a white crystalline solid. The yield is 5.1g.

The mirror image isomer excess of the title compound was higher than the compound obtained in Example 47A. (Example 47A = 86% ee, title compound > 96% ee) Analytical method for palmarity: Chiralcel OJ-H 4.6 x 250 mm, 80:20 isohexane: ethanol + 0.1% ethylenediamine, 1 ml/min, 35C, 225+-10nm after 30min.

Retention time of R image isomers = 15.91 min

Retention time of S-mirror isomer = 22.85min

^{1 H NMR (300MHz, DMSO,} 90 ℃) δ11.42 (s, 1H), 9.95 (s, 1H), 8.89 (s, 2H), 7.95 (s, 1H), 7.61-7.54 (m, 1H), 7.53-7.46(m,1H), 7.27(t, J =9.2Hz,1H), 6.94(d, J =8.5Hz,1H), 6.77(d, J =8.3Hz,1H),4.97-4.90(m , 1H), 4.30 (s, 2H), 3.74-3.59 (m, 6H), 3.35-3.02 (m, 9H), 2.93 (d, J = 14.6 Hz, 2H), 2.76-2.61 (m, 2H), 2.44 (d, J = 14.6 Hz, 2H), 2.29-2.23 (m, 2H), 2.22-2.16 (m, 2H), 2.13-2.00 (m, 2H), 1.96-1.68 (m, 6H), 1.34 ( d, J = 6.9 Hz, 6H), 1.32-1.20 (m, 4H), 1.06 (s, 6H), 0.76 (s, 6H) Two unobservable exchangeable protons.

XRPD pattern of di(1S)-(+)-10-camphorsulfonate

Deformation A

Partial characteristic peak of deformation A

Example 47C

(R)-7-(2-(2-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one fumarate variant A

(R)-7-(2-(2-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one (13 mg) (Example 47D, step A) A solution of fumaric acid (2 mg) in methanol (0.75 mL) was stirred at RT for 7 days. The white solid formed was isolated and dried under high vacuum to afford title compound as a white solid. The yield is 5 mg.

m/z 670(M+H) ⁺ (APCI)

XRPD pattern of fumarate deformation A

Some characteristic peaks of the deformed A of fumarate

Example 47D

(R)-7-(2-(2-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one monofumarate variant Ba) (R)-7-(2-(2-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one

(R)-7-(2-(2-Fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di(1S)-(+) -10-camphorsulfonate (Example 47B) (4.1 g) was dissolved between 2-methyl-THF (100 mL) and saturated sodium bicarbonate (100 mL). The layers were separated and washed with EtOAc EtOAc m. The resulting glassy solid was triturated twice with diethyl ether to give the subtitle compound as a white solid. The yield was 2.6 g.

b) (R)-7-(2-(2-Fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Decampanic-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzene well [d]thiazole-2(3H)-one monofumarate variant B

(R)-7-(2-(2-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9 -diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one (2.4g) was dissolved at 50 ° C Ethanol (240 mL). Fumaric acid (0.416 g) was added and the mixture was heated under reflux until a clear solution formed. The resulting solution was cooled to 60 ° C and inoculated with Form A (Example 47C) (50 mg). The resulting mixture was stirred overnight at 50 ° C and then allowed to cool to RT over 3 hr. The isolated white solid was filtered, washed with EtOAc (EtOAc) The yield was 2.3 g.

m/z 670(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, DMSO, 90 ° C) δ 7.89 (s, 1H), 7.19-7.09 (m, 2H), 7.01 (dd, J = 9.9, 8.3 Hz, 1H), 6.86 (d, J = 8.2 Hz, 1H), 6.69 (d, J = 8.5 Hz, 1H), 6.59 (d, J = 1.0 Hz, 2H), 4.65-4.59 (m, 1H), 3.73-3.56 (m, 6H), 3.41 (s) , 2H), 3.30 (seven peaks, J = 3.3 Hz, 1H), 2.90-2.71 (m, 6H), 2.42-2.26 (m, 4H), 1.75-1.64 (m, 2H), 1.59-1.48 (m, 2H), 1.35 (dd, J = 6.8, 1.2 Hz, 6H) + 6 unobserved exchangeables

XRPD pattern of fumarate deformation B

Partial characteristic peak of deformation B of fumarate

XRPD-PANalytical CubiX PRO

The XRPD data was collected in a θ-2θ configuration with a PANalytical CubiX PRO in a 2° to 40° 2θ scan range at 100° exposure in 0.02° increments. The X-ray system was generated from a copper elongated focus tube operating at 45 kV and 40 mA. The wavelength of copper X-ray is 1.5418 . Data was collected on a zero background holder with ~2 mg of compound placed. The holder is made of tantalum single crystal and has been cut along a non-diffractive plane and subsequently polished on an optical surface grinder. X-rays incident on this surface are eliminated by Bragg extinction.

Example 48

(R)-7-(2-(2-fluoro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-fluoro-3-(2-hydroxyethyl)benzaldehyde

2,2,6,6-Tetramethylpiperidine (10.8 mL) was added to a solution of butyllithium (1.6M in hexanes, 40.1 mL) in THF (40 mL). A solution of 2-(2-fluorophenyl)ethanol (3 g) in THF (40 mL) was added dropwise and the mixture was stirred at -70 ° C for 6 h. DMF (8.29 mL) was then added and the mixture was stirred at -70 ° C for 1 h. The mixture was then allowed to warm to RT and stirred for 70 h. The reaction was quenched with EtOAc (2M, 50 mL)EtOAc. The aqueous phase was extracted with ethyl acetate (2×100 mL). The combined organic solution was washed with EtOAc EtOAc (EtOAc m. The residue was purified by silica gel chromatography eluting with 4:1 to 2:1 isohexane:EtOAc. The fractions containing the product are combined and evaporated to give the sub-title compound as a yellow oil. The yield was 1.2g.

_{^{1 H NMR (300MHz, CDCl 3}} ) δ10.37 (s, 1H), 7.80-7.71 (m, 1H), 7.54 (td, J = 7.4,1.9Hz, 1H), 7.22 (t, J = 7.6Hz, 1H), 3.96-3.87 (m, 2H), 2.99 (td, J = 6.5, 1.2 Hz, 2H), 1.53 (t, J = 5.6 Hz, 1H).

b) (9-(2-Fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl) (2 -isopropylthiazol-4-yl)methanone

2-Fluoro-3-(2-hydroxyethyl)benzaldehyde (Example 48, step a) (0.19 g) was added to (2-isopropylthiazol-4-yl)(1-oxa-4, 9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (0.32 g) and acetic acid (0.043 mL) in N-methyl-2- In a solution of pyrrolidone (5 mL). The resulting mixture was stirred for 1 h then sodium triacetoxyborohydride (0.24 g) was added portionwise over 5 min. The resulting mixture was stirred overnight, diluted with EtOAc (20 mL) and applied to EtOAc (EtOAc) The column was washed with acetonitrile (50 mL) and dissolved in 10% ""&quot;&quot; The eluent was evaporated, azeotroped with toluene and purified by silica gel chromatography eluting with 77.5:17.5. 5 isohexane:ethyl acetate:triethylamine to 95:5 ethyl acetate:triethylamine gradient. The fractions containing the product are combined and evaporated to give the sub-title compound as a yellow gum. The yield was 0.33 g.

m/z 462(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.91 (s, 1H), 7.23 - 7.13 (m, 2H), 7.03 (t, J = 7.5 Hz, 1H), 4.40 (t, J = 5.3 Hz, 1H), 3.72-3.56 (m, 9H), 3.48 (s, 2H), 2.75 (t, J = 6.9 Hz, 2H), 2.46-2.26 (m, 4H), 1.75-1.63 (m, 2H) , 1.59-1.46 (m, 2H), 1.35 (d, J = 6.9 Hz, 6H).

c (R)-7-(2-(2-fluoro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

Add TFA (0.044 mL) to (9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 4-Base)(2-Isopropylthiazol-4-yl)methanone (Example 48, step b) (0.3 g) in DCM (5 mL) The mixture was stirred for 5 min, then Dess-Martin periodinane (0.37 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (5 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (2 mL) followed by acetic acid (0.033 mL) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.28 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.054 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 30% acetonitrile). The title compound was obtained as a white solid. The yield was 0.14 g.

m/z 670(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.27 (s, 1H), 7.94 (s, 1H), 7.51 (t, J = 6.9 Hz, 1H), 7.44 (t, J = 7.0 Hz) , 1H), 7.25 (t, J = 7.7 Hz, 1H), 6.94 (d, J = 8.2 Hz, 1H), 6.78 (d, J = 8.5 Hz, 1H), 4.93 (dd, J = 8.3, 4.7 Hz , 1H), 4.34 (s, 2H), 3.77-3.60 (m, 6H), 3.36-3.04 (m, 11H), 2.09-1.97 (m, 2H), 1.87-1.72 (m, 2H), 1.35 (d , J = 6.9 Hz, 6H). Five unobserved exchangeable protons.

Example 49

(R)-4-hydroxy-7-(1-hydroxy-2-(2-(5-((4-(2-methylbenzo[d]thiazol-5-carbonyl)-1-oxa-4) ,9-diazaspiro[5.5]undecane-9-yl)methyl)thiophen-3-yl)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one -trifluoroacetate

a) (9-((4-(2-(Third-butyldimethylsilyloxy)ethyl)thiophen-2-yl)methyl)-1-oxa-4,9-diaza Heterospiro[5.5]undecane-4-yl)(2-methylbenzo[d]thiazol-5-yl)methanone

The HATU (0.333g) was added to 2-methyl-benzo [d] thiazole-5-carboxylic acid (0.17g), 9 - (( 4- (2- ( tertiary - butyl dimethyl silicone alkyloxy) Ethyl)thiophen-2-yl)methyl)-1-oxa-4,9-diazaspiro[5.5]undecane (Example 43, step b) (0.3 g) and triethylamine ( 0.407 mL) in a stirred solution in DMF (3 mL). After 1 h, the reaction mixture was partitioned between water and ethyl acetate. The ethyl acetate layer was washed twice with water, brine brine and dried over magnesium sulfate. Purification by silica gel chromatography eluting with ethyl acetate: hexanes: triethylamine 2: 8:1 to give the subtitle compound. The yield was 0.36 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.06 (d, J = 8.3 Hz, 1H), 7.38 (d, J = 8.3 Hz, 1H), 7.78 (s, 1H), 6.96 (s) , 1H), 6.78 (s, 1H), 3.77 (t, J = 6.7 Hz, 2H), 3.68-3.63 (m, 2H), 3.58 (s, 2H), 3.53-3.46 (m, 2H), 3.42 3.36 (m, 2H), 2.82 (s, 3H), 2.70 (t, J = 6.8 Hz, 2H), 2.42-2.31 (m, 4H), 1.81-1.72 (m, 2H), 1.52-1.41 (m, 2H), 0.86 (s, 9H), 0.00 (s, 6H).

b) (9-((4-(2-hydroxyethyl))thiophen-2-yl)methyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl (2-methylbenzo[d]thiazol-5-yl)methanone

The TBAF (1M solution in THF, 1.5mL) was added to (9 - ((4- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethyl) thiophen-2-yl) methyl) 1-oxathia-4,9-diazaspiro[5.5]undecane-4-yl)(2-methylbenzo[d]thiazol-5-yl)methanone (Example 49, step a) (0.36 g) in a stirred solution in THF (2 mL). After 1 h, the reaction evaporated to a gum. Purification by silica gel chromatography, eluting with ethyl acetate: triethylamine 20:1 affords the subtitle compound. The yield was 0.2 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.05 (d, J = 7.5 Hz, 1H), 7.37 (d, J = 7.5 Hz, 1H), 7.86 (s, 1H), 6.94 (s) , 1H), 6.76 (s, 1H), 4.30-4.23 (m, 1H), 3.68-3.54 (m, 6H), 3.54-3.44 (m, 2H), 3.43-3.33 (m, 2H), 2.81 (s , 3H), 2.71-2.62 (m, 2H), 2.47-2.29 (m, 4H), 1.81-1.71 (m, 2H), 1.55-1.40 (m, 2H).

c) 2-(5-((4-(2-methylbenzo[d]thiazol-5-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)methyl)thiophen-3-yl)acetaldehyde

Add Dess-Martin periodinane (0.27g) to (9-((4-(2-hydroxyethyl))thiophen-2-yl)methyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-4-yl)(2-methylbenzo[d]thiazol-5-yl)methanone (Example 49, step b) (0.20 g) and trifluoroacetic acid (0.033 mL) ) in a stirred solution in DCM (5 mL). After 40 min, the reaction mixture was crystallisjjjjjjjjjjjjjjjjjjj The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. The yield was 0.2 g. Used directly in the next steps.

m/z 470(M+H) ⁺ (APCI)

d) (R)-4-hydroxy-7-(1-hydroxy-2-(2-(5-((4-(2-methylbenzo[d]thiazol-5-carbonyl)-1-oxa) -4,9-diazaspiro[5.5]undecane-9-yl)methyl)thiophen-3-yl)ethylamino)ethyl)benzo[d]thiazole-2(3H)- Keto di-trifluoroacetate

Acetic acid (0.037 mL) was added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.168 g) and 2-(5-((4-(2-methylbenzo[d]thiazol-5-carbonyl)-1-oxa-4,9-diaza A stirred solution of spiro[5.5]undecane-9-yl)methyl)thiophen-3-yl)acetaldehyde (Example 49, step c) (0.200 g) in MeOH (8 mL). After 1 min, sodium cyanoborohydride (0.107 g) was added. After 3h, the reaction mixture was filtered and purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in acetonitrile 10 to 35%) by. The fractions containing the pure product were combined and evaporated to dryness. The title compound was obtained as a white solid. The yield was 0.15 g.

m/z 680(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.29 (s, 1H), 8.07 (d, J = 8.2 Hz, 1H), 7.90 (s, 1H), 7.42 - 7.37 (m, 2H) , 7.15 (s, 1H), 6.93 (d, J = 8.7 Hz, 1H), 6.77 (d, J = 8.7 Hz, 1H), 4.94 - 4.86 (m, 1H), 4.50 - 4.41 (m, 2H), 3.73-3.63 (m, 2H), 3.58-3.33 (m, 4H), 3.29-2.90 (m, 10H), 2.81 (s, 3H), 2.16-2.00 (m, 2H), 1.78-1.62 (m, 2H) ). 4 unobserved exchangeable protons.

Example 50

(R)-7-(2-(3-((4-(2-)-tert-butylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecene Alkan-9-yl)methyl)-5-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (2-t-butylthiazol-4-yl)(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro [5.5] undecane-4-yl)methanone

2,2,2-trifluoro-1-(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] A solution of the carboxane-4-yl)ethanone (Example 41, step d) (0.21 g) in MeOH (3 mL)EtOAc. The mixture was evaporated to dryness under reduced pressure. The residue was dissolved in DMF (7mL) and with a third 2- - butyl-4-carboxylic acid (0.096 g), followed by triethylamine (0.290 mL), and followed by HATU (0.257g) processing, and forming the mixture Stir at 20 ° C for 1 hour. The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The yield was 0.22 g.

m/z 476(M+H) ⁺ (APCI)

b) (R)-7-(2-(3-((4-(2-)-tert-butylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)-5-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetic acid salt

(2-tertiary - butyl-thiazol-4-yl) (9- (3-fluoro-5- (2-hydroxyethyl) benzyl) -l-oxa-4,9-diazaspiro [5.5 a solution of the undecane-methyl ketone (Example 50, step a) (0.22 g) in DCM (20 mL) eluting with trifluoroacetic acid (0.036 mL) The alkane treatment (0.255 g) and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.026 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.182 g) and acetic acid (0.026 mL) in methanol (15 mL). The mixture was cooled in an ice-bath and treated with sodium cyanoborohydride (0.058 g). The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.135 g.

m/z684(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.95 (s, 1H), 7.27-7.16 (m, 3H), 6.93 (d, J = 8.5 Hz, 1H), 6.77 (d, J = 8.5 Hz, 1H), 4.94-4.88 (m, 1H), 4.21 (s, 2H), 3.71 (s, 4H), 3.66 (s, 2H), 3.26 (t, J = 7.9 Hz, 2H), 3.15- 2.98 (m, 8H), 2.06-1.95 (m, 2H), 1.79-1.67 (m, 2H), 1.41 (s, 9H). Six unobserved exchangeable protons.

Example 51

(R)-5-(2-(2-(5-((2,2-difluoro-4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diaza) Heterospiro[5.5]undecane-9-yl)methyl)thiophen-3-yl)ethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di- Trifluoroacetate

a) (9-((4-(2-(Third-butyldimethylsilyloxy)ethyl)thiophen-2-yl)methyl)-2,2-difluoro-1-oxa -4,9-diazaspiro[5.5]undecane-4-yl)(2-isopropylthiazol-4-yl)methanone

(2,2-Difluoro-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-isopropylthiazol-4-yl)methanonetrifluoro acetate (Example 44, step b) (0.40g) was added to 4- (2- (third - butyldimethylsilyl silicon alkyloxy) ethyl) thiophene-2-carbaldehyde and 3- (2- ( third - butyldimethyl silicon alkyloxy) ethyl) thiophene-2-carbaldehyde of 4: 1 mixture (Example 27, step b) (0.36g) and AcOH (0.05mL) in N- methyl - Stirred solution in 2-pyrrolidone (5 mL). After 5 min, sodium triethoxysulfonium borohydride (0.28 g) was added. After 16 h, water was added and the mixture was extracted with ethyl acetate. The ethyl acetate layer was washed three times with water and evaporated in vacuo. Purification by silica gel chromatography, eluting with 20:80:5 ethyl acetate: isohexane: triethylamine, and separating the two isomeric products to give a sub-title compound. The yield was 0.24 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.11 (s, 1H), 7.00 (s, 1H), 6.82 (s, 1H), 4.28 - 4.16 (m, 2H), 4.05 - 3.95 ( m, 2H), 3.78 (t, J = 6.6 Hz, 2H), 3.64 (s, 2H), 3.40-3.31 (m, 1H), 2.71 (t, J = 6.6 Hz, 2H), 1.85-1.77 (m , 2H), 1.74-1.64 (m, 2H), 1.39 (d, J = 6.8 Hz, 6H), 0.86 (s, 9H), 0.86 (s, 6H), 4 protons under solvent peaks.

b) 2,2-difluoro-9-((4-(2-hydroxyethyl)thiophen-2-yl)methyl)-1-oxa-4,9-diazaspiro[5.5] eleven Cycloalkyl-4-yl)(2-isopropylthiazol-4-yl)methanone

The TBAF (in THF 1M, 1.5mL) was added to (9 - ((4- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethyl) thiophen-2-yl) methyl) - 2,2-Difluoro-1-oxa-4,9-diazaspiro[5.5]undecethane-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 51 Step a) (0.30 g) in a stirred solution in THF (3 mL). After 1 h, the reaction evaporated to a gum in vacuo. Purification by silica gel chromatography, eluting with ethyl acetate: triethylamine 20:1 affords the subtitle compound. The yield was 0.2 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 8.19 (s, 1H), 7.02 (s, 1H), 6.81 (s, 1H), 4.60 (t, J = 5.3 Hz, 1H), 4.54-4.39 ( m, 1H), 4.20-3.98 (m, 3H), 3.88-3.75 (m, 1H), 3.66-3.53 (m, 5H), 2.65 (t, J = 7.0 Hz, 3H), 1.81-1.73 (m, 2H), 1.72-1.60 (m, 2H), 1.36 (d, J = 6.4 Hz, 6H). 2 protons blurred by solvent peaks.

c) 2-(5-((2,2-difluoro-4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undec carbon Alkan-9-yl)methyl)thiophen-3-yl)acetaldehyde

Add Dess-Martin periodinane (0.20 g) to (2,2-difluoro-9-((4-(2-hydroxyethyl)thiophen-2-yl)methyl)-1-oxa- 4,9-diazaspiro[5.5]undecyl-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 51, step b) (0.15 g) and trifluoroacetic acid (0.031 mL) in a stirred solution in DCM (4 mL). After 40 min, the reaction mixture was crystallisjjjjjjjjjjjjjjjjjjj The mixture was extracted with EtOAc (EtOAc) EtOAc (EtOAc) The yield was 0.19 g. Use directly.

m/z 484(M+H) ⁺ (APCI)

d) (R)-5-(2-(2-(5-((2,2-difluoro-4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9- Diazaspiro[5.5]undecane-9-yl)methyl)thiophen-3-yl)ethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one Di-trifluoroacetate

The (R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8-hydroxy-quinolin -2 (1H) - one (WO 2004106333) ( 0.156 g) was added to 2-(5-((2,2-difluoro-4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] To a stirred solution of the undecane-9-yl)methyl)thiophen-3-yl)acetaldehyde (Example 51, step c) (0.15 g) and EtOAc (EtOAc) After 5 minutes, sodium cyanoborohydride (0.078 g) was added. After 3 h, the reaction mixture was evaporated in vacuo tolululululu The ethyl acetate solution was washed with brine, dried over sodium sulfate, filtered and evaporated in vacuo. The resulting gum was dissolved in THF (2 mL) and triethylamine trihydrofluoric acid salt (0.064 g). After 16 h, toluene was added and the solution was evaporated in vacuo. Purification by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 35% acetonitrile). The title compound was combined to give the title compound as a solid. The yield was 0.14 g.

m/z 688(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ 10.52-10.47 (m, 2H), 8.85-8.70 (m, 2H), 8.22 (s, 1H), 8.15 (d, J = 9.6Hz, 1H), 7.49 -7.43 (m, 1H), 7.21 - 7.12 (m, 1H), 7.15 (d, J = 6.9 Hz, 1H), 6.99 (d, J = 8.1 Hz, 1H), 6.58 (d, J = 10.6 Hz, 1H), 6.24-6.17 (m, 1H), 5.33 (d, J = 8.9 Hz, 1H), 4.66-4.51 (m, 2H), 3.44-2.91 (m, 10H), 2.15-2.03 (m, 2H) , 1.95-1.77 (m, 2H), 1.35 (d, J = 6.9 Hz, 6H). 6 protons blurred by solvent peaks.

Example 52

(R)-7-(2-(3-chloro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(3-(bromomethyl)-5-chlorophenyl)acetic acid

Benzoyl peroxide (0.112 g) was added to 2-(3-chloro-5-methylphenyl)acetic acid (0.752 g) and N-bromosuccinimide (0.801 g) in DCM (15 mL) The suspension was formed and the resulting mixture was heated overnight at 50 ° C under nitrogen. The mixture was concentrated in vacuo to remove dichloromethane. The solution was heated at 85 ° C for 4 hours under nitrogen and then cooled. The solution was washed three times with water and once with brine, then dried over anhydrous MgSO.sub. Light yellow solid. The yield was 0.735 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.33 - 7.31 (m, 1H), 7.24 - 7.22 (m, 1H), 7.21-7.18 (m, 1H), 4.41 (s, 2H), 3.64 (s, 2H) ). An unobserved exchangeable proton.

b ) 2-(3-(bromomethyl)-5-chlorophenyl)ethanol

Borane-methylthio complex solution (2M in THF, 2.8 mL) was added portionwise to 2-(3-(bromomethyl)-5-chlorophenyl)acetic acid at room temperature over 5 min. Example 52, step a) (0.73 g) in EtOAc (EtOAc) The resulting effervescent solution was stirred for 1 hour, then cooled in ice-water, and the reaction was quenched (5 mL) over 5 min. The solution was stirred at room temperature for an additional 20 minutes and then concentrated in vacuo. The residue was purified by flash chromatography eluting with EtOAc EtOAc The yield was 0.46 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.29-7.24 (m, 1H), 7.19-7.13 (m, 2H), 4.41 (s, 2H), 3.87 (t, J = 6.5Hz, 2H), 2.84 ( t, J = 6.4 Hz, 2H). An unobserved exchangeable proton.

c) (9-(3-Chloro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -isopropylthiazol-4-yl)methanone

Add triethylamine (0.18 mL) to (2-isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undec-4-yl)methanone Trifluoroacetate (Example 22, step b) (0.217 g) and 2-(3-(bromomethyl)-5-chlorophenyl)ethanol (Example 52, step b) (0.160 g) in acetonitrile The suspension in 5 mL) was stirred and the solution formed was stirred overnight at room temperature. The solution was then purified by flash chromatography on ruthenium dioxide eluting with 1:5:94 triethylamine:methanol:dichloromethane to give a gum. The gum was dissolved in dichloromethane and the solution was washed with EtOAc EtOAc EtOAc EtOAc The yield was 0.254 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ7.90 (s, 1H), 7.21-6.92 (m, 3H), 4.31 (t, J = 5.3Hz, 1H), 3.79-3.49 (m, 8H), 3.42 (s, 2H), 3.31 (seven peak, J = 6.8 Hz, 1H), 2.71 (t, J = 6.7 Hz, 2H), 2.43 - 2.22 (m, 4H), 1.77-1.65 (m, 2H), 1.60-1.48 (m, 2H), 1.36 (d, J = 6.9 Hz, 6H).

d) 2-(3-chloro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)methyl)phenyl)acetaldehyde

(9-(3-Chloro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-iso A solution of the propyl thiazol-4-yl)methanone (Example 52, step c) (0.242 g) in EtOAc (EtOAc)EtOAc. Dess-Martin periodinane (0.323 g) was added, after which the mixture was taken out from the cooling bath and stirred at room temperature for 30 minutes. The solution was diluted with a saturated aqueous solution of sodium thiosulfate (5 mL), saturated sodium bicarbonate (5 mL) and ethyl acetate (5 mL), and the mixture was stirred vigorously for 10 min. The mixture was then extracted with EtOAc (EtOAc)EtOAc. The yield was 0.297 g.

m/z 476(M+H) ⁺ (APCI)

e) (R)-7-(2-(3-chloro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

( R )-7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) A solution of 0.220 g) in MeOH (3 mL) wasEtOAc (EtOAc) Subsequent addition of 2-(3-chloro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 a solution of -methyl)phenyl)acetaldehyde (Example 52, step d) (0.296 g) in MeOH (4 mL), and the resulting mixture was stirred at room temperature for 5 min then cooled in ice-water. Treatment with sodium triethoxysulfonate (0.181 g). The mixture was stirred in ice-water for 10 min and then at room temperature over 45 min then cooled in ice-water and taken over <RTI ID=0.0> The mixture was stirred overnight and allowed to slowly warm to room temperature. The next morning, the mixture was concentrated in vacuo. The residue was dissolved in a mixture of EtOAc (EtOAc ⁾ (EtOAc ⁾ (EtOAc) The product containing fractions were concentrated in vacuo and co-evaporated three times from acetonitrile to give a colourless residue. The residue was triturated with EtOAc (EtOAc)EtOAc. The yield was 0.092 g.

m/z 686(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ 7.94 (s, 1H), 7.50-7.44 (m, 1H), 7.44-7.37 (m, 1H), 7.34-7.26 (m, 1H), 6.93 (d, J = 8.5 Hz, 1H), 6.77 (d, J = 8.2 Hz, 1H), 4.90 (dd, J = 7.9, 5.4 Hz, 1H), 4.26 - 4.08 (m, 2H), 3.77 - 3.59 ( m, 6H), 3.35-3.21 (m, 3H), 3.16-2.91 (m, 8H), 2.06-1.90 (m, 2H), 1.81-1.63 (m, 2H), 1.35 (d, J = 6.9 Hz, 6H). Six unobserved exchangeable protons.

Example 53

(R)-7-(2-(3-fluoro-5-((4-(4-isopropylthiazol-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzene well [d]thiazole-2(3H)-one di-trifluoroacetate

a) 4-isopropylthiazole-2-carboxylic acid ethyl ester

1-Bromo-3-methylbutan-2-one (3.72 g) in ethanol (15 mL) was added dropwise to ethyl 2-amino-2-ylthioacetate (3 g) in ethanol over 15 min. (120 mL) in a refluxing solution. The mixture was heated under reflux for 2 hours and then cooled to room temperature. The solvent was distilled under reduced pressure to a volume of 30 mL, this solution was added to ice/water (200 mL), and the mixture was neutralized by dropwise addition of a '880' concentrated aqueous ammonia solution. The mixture was extracted with EtOAc (EtOAc)EtOAc. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 2.2 g.

m/z 200(M+H) ⁺ (APCI)

b) 4-isopropylthiazole-2-carboxylic acid

A solution of ethyl 4-isopropylthiazole-2-carboxylate (Example 53, step a) (2.2 g) in a mixture of methanol (10 mL) and THF (20 mL) with lithium hydroxide (0.264 g) Solution in 20 mL). The reaction mixture was stirred at 20 ° C for 24 hours. The organic solvent was removed under reduced pressure and the remaining aqueous mixture was partitioned between ethyl acetate and water. The aqueous layer was acidified with EtOAc (EtOAc)EtOAc. The crude product was triturated with cyclohexane to give the subtitle compound. The yield was 0.93 g.

m/z 170(MH) ^- (APCI)

¹ H NMR (400 MHz, D _{6 -} DMSO) δ 7.67 (s, 1H), 3.16-3.05 (m, 1H), 1.26 (d, 6H). An unobserved exchangeable proton.

c) (9-(3-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(4 -isopropylthiazol-2-yl)methanone

2,2,2-trifluoro-1-(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] A solution of the carboxane-4-yl)ethanone (Example 41, step d) (0.21 g) in MeOH (3 mL)EtOAc. The mixture was evaporated to dryness under reduced pressure. The residue was dissolved in DMF (7 mL) EtOAc (EtOAc (EtOAc) Treated and the resulting mixture was stirred at 20 ° C for 1 hour. The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The yield was 0.325 g.

m/Z462(M+H) ⁺ (APCI)

d) (R)-7-(2-(3-Fluoro-5-((4-(4-isopropylthiazol-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

(9-(3-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(4-iso A solution of propyl thiazol-2-yl)methanone (Example 53, step c) (0.23 g) in DCM (20 mL (0.275 g) and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.029 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.196 g) and acetic acid (0.029 mL) in methanol (15 mL). The mixture was cooled in an ice-bath and treated with sodium cyanoborohydride (0.063 g). The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.140 g.

m/z 670(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.27 (s, 1H), 7.51 (s, 1H), 7.28-7.16 (m, 3H), 6.93 (d, J = 8.2 Hz, 1H) , 6.77 (d, J = 8.2 Hz, 1H), 4.95-4.88 (m, 1H), 4.26 (s, 2H), 4.19-3.82 (m, 4H), 3.80-3.74 (m, 2H), 3.27 (t , J = 7.9 Hz, 2H), 3.19-2.98 (m, 9H), 2.09-1.98 (m, 2H), 1.85-1.73 (m, 2H), 1.27 (d, J ⁼ 7.1 Hz, 6H). Five unobserved exchangeable protons.

Example 54

(R)-7-(2-(3-fluoro-5-((4-(5-isopropylthiazol-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 5-isopropylthiazole-2-carboxylic acid ethyl ester

A solution of bromine (0.162 mL) in a mixture of DCM (1. <RTI ID=0.0></RTI> </RTI> <RTI ID=0.0> (0 mL) in a mixture of 0 ° C in a cooled solution. The mixture was stirred at 5 ° C for 2 hours. The reaction mixture was cooled to 0 ° C and the solvent was removed under a stream of nitrogen. The residue, still at 0&lt;0&gt;C, was treated in portions with ethyl 2-amino-2-sylthioacetate (0.42 g). The resulting mixture was heated from 0 ° C to 70 ° C under nitrogen for 15 minutes. At the end of this time, the mixture was cooled with EtOAc EtOAc m. The crude product was purified by flash chromatography on silica gel eluting with 30% ethyl acetate in isohexane. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.105 g.

m/z 200(M+H) ⁺ (APCI)

b) 5-isopropylthiazole-2-carboxylic acid

A solution of ethyl 5-isopropylthiazole-2-carboxylate (Example 54, step a) (0.105 g) in MeOH (5 mL) And the resulting mixture was vigorously stirred at 20 ° C for 2 hours. Methanol was distilled off under reduced pressure and the residual aqueous solution was diluted with brine (10 mL). The aqueous layer was washed with diethyl ether, then cooled in an ice-bath, and acidified by dropwise addition of concentrated aqueous hydrochloric acid. The mixture was extracted twice with ethyl acetate. The combined ethyl acetate layer was washed with brine, dried over sodium sulfate, filtered and evaporated The yield was 0.048 g.

¹ H NMR (400 MHz, D _{6 -} DMSO) δ 7.84 (s, 1H), </ RTI></RTI></RTI></RTI></RTI></RTI><RTIgt; An unobserved exchangeable proton.

c) (9-(3-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl) (5 -isopropylthiazol-2-yl)methanone

2,2,2-trifluoro-1-(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] A solution of the carnamid-4-yl)ethanone (Example 41, step d) (0.113 g) in MeOH (3 mL)EtOAc. The mixture was evaporated to dryness under reduced pressure. The residue was dissolved in DMF (7 mL) EtOAc (EtOAc (EtOAc) Treated and the resulting mixture was stirred at 20 ° C for 1 hour. The mixture was partitioned between EtOAc and EtOAc. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 104 mg.

m/z 462(M+H) ⁺ (APCI)

d) (R)-7-(2-(3-Fluoro-5-((4-(5-isopropylthiazol-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

(9-(3-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(5-iso A solution of the propyl thiazol-2-yl)methanone (Example 54, step c) (0.104 g) in DCM (20 mL (0.124 g) and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.013 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.089 g) and acetic acid (0.013 mL) in methanol (15 mL). The mixture was cooled in an ice-bath and treated with sodium <RTI ID=0.0> The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.045 mg.

m/z 670(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ7.67 (s, 1H), 7.28-7.16 (m, 3H), 6.93 (d, J = 8.5Hz, 1H), 6.77 (d, J = 8.5 Hz, 1H), 4.94-4.89 (m, 1H), 4.26 (s, 2H), 4.11-3.79 (m, 4H), 3.78-3.73 (m, 2H), 3.31-3.24 (m, 2H), 3.18 -2.99 (m, 9H), 2.06-1.97 (m, 2H), 1.85-1.74 (m, 2H), 1.31 (d, J = 7.0 Hz, 6H). Six unobserved exchangeable protons.

Example 55 (R)-7-(2-(3-fluoro-5-((4-(2-isopropylthiazol-5-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-isopropylthiazole-5-carboxylic acid ethyl ester

A mixture of 2-methylpropanethioguanamine (3.3 g) and ethyl 2-chloro-3-oxopropionate (4.82 g) in acetone (50 mL) was evaporated. The mixture was cooled to room temperature and the solvent was removed under reduced pressure. The crude product was purified by flash chromatography on silicon dioxide using 12% isohexane in ethyl acetate as the ^solvent. The pure fractions were evaporated to dryness to give the subtitle compound. The yield is 1.6g.

m/z 200(M+H) ⁺ (APCI)

b) 2-isopropylthiazole-5-carboxylic acid

A solution of 2-isopropylthiazol-5-carboxylate (Example 55, step a) (0.33 g) in MeOH (EtOAc) (EtOAc) The resulting mixture was stirred at 20 ° C for 2 hours. Methanol was evaporated under reduced pressure and the residue was partitioned between diethyl ether and brine. The aqueous layer was acidified by dropwise addition of dilute hydrochloric acid and extracted with ethyl acetate. The organic layer was washed with EtOAc (EtOAc m. The yield was 0.185 g.

m/z 172(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D6-DMSO) δ 13.38 (s, 1H), 8.22 (s, 1H), 3.35-3.26 (m, 1H), 1.34 (d, J = 6.8 Hz, 6H).

c) 2-(3-(1-oxa-4,9-diazaspiro[5.5]undecal-9-ylmethyl)-5-fluorophenyl)ethanol

Add '880' ammonia solution (5 mL) to 2,2,2-trifluoro-1-(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4, 9-Diazaspiro[5.5]undecane-4-yl)ethanone (Example 41, step d) (2.2 g) in MeOH (25 mL). The resulting mixture was stirred for 90 min and the solvent was evaporated. The residue was azeotroped three times with acetonitrile and concentrated to give a crude title compound. The yield was 1.96 g.

m/z 309(M+H) ⁺ (APCI)

_{^{1 H NMR (300MHz, D 6}} -DMSO) δ6.97 (s, 1H), 6.94 (s, 1H), 6.91 (s, 1H), 3.65-3.56 (m, 4H), 3.49-3.39 (m, 2H ), 2.85-2.78 (m, 2H), 2.76-2.68 (m, 4H), 2.48-2.39 (m, 2H), 2.32-2.21 (m, 2H), 1.89-1.77 (m, 2H), 1.58-1.44 (m, 2H). Two unobserved exchangeable protons.

d) (9-(3-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -isopropylthiazol-5-yl)methanone

2-(3-(1-oxa-4,9-diazaspiro[5.5]undecal-9-ylmethyl)-5-fluorophenyl)ethanol (Example 55, step c) A solution of 0.153 g) in DMF (7 mL) was obtained eluted elute The mixture was cooled to 0 ° C and HATU (0.245 g) was added. The reaction mixture was stirred at 20 ° C for 2 hours. The mixture was partitioned between EtOAc and EtOAc. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.130 g.

m/z 462(M+H) ⁺ (APCI)

e) (R)-7-(2-(3-Fluoro-5-((4-(2-isopropylthiazol-5-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

(9-(3-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-iso A solution of propylthiazole-5-yl)methanone (Example 55, step d) (0.13 g) in DCM (20 mL) (0.155 g) and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.016 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.111 g) and acetic acid (0.016 mL) in methanol (15 mL). The mixture was cooled in an ice-bath and treated with sodium br The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield is 0.1 g.

m/z 670(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.27 (s, 1H), 7.92 (s, 1H), 7.30-7.18 (m, 3H), 6.93 (d, J = 8.2Hz, 1H) , 6.77 (d, J = 8.2 Hz, 1H), 4.95-4.89 (m, 1H), 4.30 (s, 2H), 3.75-3.70 (m, 2H), 3.68-3.63 (m, 2H), 3.54 (s) , 2H), 3.33 - 3.24 (m, 3H), 3.22-3.00 (m, 8H), 2.07-1.98 (m, 2H), 1.84-1.72 (m, 2H), 1.35 (d, J = 6.9 Hz, 6H ). Five unobserved exchangeable protons.

Example 56

(R)-7-(2-(3-fluoro-5-((4-(2-propylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] eleven Cycloalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-butylammonium-3-mercaptopropionate ethyl ester

To a suspension of 2-amino-3-mercaptopropionic acid ethyl ester hydrochloride (5 g) in DCM (50 mL). The resulting mixture was cooled in an ice/salt bath and butyl chloride (3.3 mL) was added dropwise. The resulting mixture was allowed to warm to RT and stirred overnight. The reaction was quenched with saturated sodium bicarbonate solution (50 mL) and partition. The aqueous phase was extracted with DCM (2×100 mL). The combined organic solution was washed with brine (100 mL) dried over sodium sulfate. The white solid formed was purified by chromatography on silica gel eluting with 4:1 to 1:1 isohexane:ethyl acetate gradient. The fractions containing the product are combined and evaporated to give the subtitle compound as a white solid. The yield is 3g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 8.21 (d, J = 7.7 Hz, 1H), 4.43-4.37 (m, 1H), 4.13-4.06 (m, 2H), 2.88-2.80 (m, 1H) ), 2.78-2.69 (m, 1H), 2.12 (t, J = 7.2 Hz, 2H), 1.60-1.47 (m, 2H), 1.22-1.14 (m, 3H), 0.90-0.81 (m, 3H). An unobserved exchangeable proton.

m/z 218(M-H)-(APCI)

b) Ethyl 2-propylthiazole-4-carboxylate

Add HCl solution (4M in 1,4-dioxane, 20 mL) to ethyl 2-butyiamine-3-mercaptopropanoate (Example 56, step a) (3 g) in ethanol (20 mL) The solution was formed and the mixture formed was stirred overnight. The solvent was evaporated and the residue was azeotroped twice with toluene. The residue was redissolved in acetonitrile (50 mL). EtOAc (11.9 g). The reaction was filtered through a pad of Celite. The filter pad was washed with MeCN (3 x 100 mL). The combined filtrate and washings were evaporated and the residue was purified by chromatography eluting with EtOAc EtOAc. The fractions containing the product are combined and evaporated to give the clear oil of the sub-title compound. The yield was 0.24 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.05 (s, 1H), 4.42 (q, J = 7.1 Hz, 2H), 3.04 (t, J = 7.7 Hz, 2H), 1.89-1.77 (m, 2H) , 1.40 (t, J = 7.0 Hz, 3H), 1.02 (t, J = 7.4 Hz, 3H).

c) 2-propylthiazole-4-carboxylic acid

Add lithium hydroxide monohydrate (0.2 g) to a mixture of 2-propylthiazole-4-carboxylate (Example 56, step b) (0.24 g) in THF (4 mL) and water (1 mL) In solution. The resulting suspension was stirred overnight at RT. The reaction was acidified (2M, 3 mL) elut The residue was dissolved between brine (5 mL) and ethyl acetate (20 mL). The layers were separated and the aqueous extracted with EtOAc (2×20 mL). The combined organic solution was dried with sodium sulfate, filtered and evaporated elut The yield was 0.17 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 12.91 (s, 1H), 8.31 (s, 1H), 2.96 (t, J = 7.4 Hz, 2H), 1.79-1.68 (m, 2H), 0.95 (t) , J = 7.4 Hz, 3H).

d) (9-(3-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -propylthiazole-4-yl)methanone

Adding O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.25 g) to 2-(3-(1- Oxa-4,9-diazaspiro[5.5]undecal-9-ylmethyl)-5-fluorophenyl)ethanol (Example 55, step c) (0.16 g), 2-propyl A solution of thiazole-4-carboxylic acid (Example 56, step c) (0.09 g) and triethylamine (0.29 mL) in EtOAc (EtOAc) The mixture was partitioned between ethyl acetate (100 mL) and brine (100 mL). The organic phase was separated, washed with brine (2×100 mL) dried over sodium sulfate. The resulting gum was purified by silica gel chromatography eluting with 47.5:47.5:5 isohexane:ethyl acetate:triethylamine to 95:5 ethyl acetate:triethylamine. To the residue containing the product, toluene (200 mL) was added, and the mixture was evaporated to give a sub-title compound. The yield was 0.14 g.

m/z 462(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.89 (s, 1H), 6.95 (s, 1H), 6.89 (s, 1H), 6.86 (s, 1H), 4.31 (t, J = 4.7 Hz, 1H), 3.69-3.59 (m, 8H), 3.44 (s, 2H), 2.98 (t, J = 7.2 Hz, 2H), 2.72 (t, J = 6.8 Hz, 2H), 2.43-2.28 (m , 4H), 1.82-1.66 (m, 4H), 1.59-1.50 (m, 2H), 0.97 (t, J = 7.3 Hz, 3H).

e) (R)-7-(2-(3-Fluoro-5-((4-(2-propylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

Adding TFA (0.023 mL) to DCM (5 mL) (0-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diaza snail at 0 °C [5.5] undecane-4-yl)(2-propylthiazol-4-yl)methanone (Example 56, step d) (0.14 g). The mixture was stirred for 5 min, then Dess-Martin periodinane (0.19 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (5mL), followed by addition of acetic acid (0.017mL), and (R) -7- (2- amino-1-hydroxyethyl) -4-hydroxyphenyl and [d] thiazol -2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.12 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.029 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 30% acetonitrile). The title compound was obtained as a white solid. The yield was 0.077 g.

m/z 670(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.27 (s, 1H), 7.93 (s, 1H), 7.29-7.15 (m, 3H), 6.93 (d, J = 8.2 Hz, 1H) , 6.77 (d, J = 8.5 Hz, 1H), 4.95 - 4.88 (m, 1H), 4.28 - 4.21 (m, 2H), 3.73 - 3.61 (m, 6H), 3.27 (t, J = 7.9 Hz, 2H) ), 3.16-2.93 (m, 10H), 2.05-1.95 (m, 2H), 1.83-1.70 (m, 4H), 0.96 (t, J = 7.3 Hz, 3H). Five unobserved exchangeable protons.

Example 57

(R)-7-(2-(3-((4-(2-cyclopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 9-yl)methyl)-5-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzene well [d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(cyclopropanecarbamido)-3-mercaptopropionic acid ethyl ester

Add cyclopropanecarbonyl chloride (3.1 mL) to a suspension of triethylamine (4.68 mL) and ethyl 2-amino-3-mercaptopropionate hydrochloride (5.2 g) in DCM (200 mL) . The resulting mixture was allowed to warm to RT and stirred overnight. The reaction was quenched with ethanol and the solvent was evaporated. The residue was suspended in EtOAc (EtOAc) (EtOAc) (EtOAcjjjjjjj . The resulting white solid was purified by EtOAc (EtOAc) elute The yield is 5g.

_{^{1 H NMR (300MHz, D 6}} -DMSO) δ8.51 (d, J = 7.7Hz, 1H), 4.44 (td, J = 7.5,5.3Hz, 1H), 4.17-4.05 (m, 2H), 2.91- 2.67 (m, 2H), 2.53-2.52 (m, 1H), 1.78-1.60 (m, 1H), 1.19 (t, J = 6.8 Hz, 3H), 0.71 - 0.66 (m, 4H).

b) 2-cyclopropylthiazole-4-carboxylic acid ethyl ester

A solution of 2-(cyclopropanecarbamimidino)-3-mercaptopropionic acid ethyl ester (Example 57, step a) (5 g) and p-toluenesulfonic acid (0.875 g) in toluene (100 mL) Heated under Dean and Stark overnight. The reaction was cooled and the tolu~~~~~~~~~~~~ The resulting brown gum was redissolved in acetonitrile (100 mL). Manganese dioxide (40 g) was added and the mixture was heated under reflux overnight. The reaction was filtered through celite and the pad was washed with MeCN (3×100 mL). The combined filtrate and washings were evaporated and the residue was purified by chromatography eluting with EtOAc EtOAc EtOAc The fractions containing the product are combined and evaporated to give the clear oil of the sub-title compound. The yield was 0.15 g.

_{^{1 H NMR (400MHz, CDC1 3}} ) δ7.94 (s, 1H), 4.40 (q, J = 7.1Hz, 2H), 2.45-2.37 (m, 1H), 1.39 (t, J = 7.2Hz, 3H) , 1.20 - 1.04 (m, 4H).

c) 2-cyclopropylthiazole-4-carboxylic acid

Add lithium hydroxide monohydrate (0.13 g) to a mixture of 2-cyclopropylthiazole-4-carboxylate (Example 57, step b) (0.15 g) in THF (8 mL) and water (2 mL) The solution was formed and the mixture formed was stirred overnight. The reaction was acidified with aqueous HCl (2M, 3 mL) and evaporated. The residue was dissolved between brine (20 mL) and ethyl acetate (30 mL). The layers were separated and the aqueous extracted with ethyl acetate (2×30 mL). The combined organic solution was dried with sodium sulfate, filtered and evaporated elut The yield was 0.12 g.

¹ H NMR (300 MHz, D ₆ -DMSO) δ 12.88 (s, 1H), 8.19 (s, 1H), 2.48-2.37 (m, 1H), 1.18-1.09 (m, 2H), 1.01-0.94 (m) , 2H).

d) (2-propylpropylthiazol-4-yl)(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-4-yl)methanone

Adding O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.34 g) to 2-(3-(1- Oxa-4,9-diazaspiro[5.5]undecal-9-ylmethyl)-5-fluorophenyl)ethanol (Example 55, step c) (0.21 g), 2-cyclopropyl A solution of the thiazole-4-carboxylic acid (Example 57, step c) (0.12 g) and triethylamine (0.38 mL) in DMF (7 mL) The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The resulting gum was purified by silica gel chromatography eluting with 47.5:47.5:5 isohexane:ethyl acetate:triethylamine to 95:5 ethyl acetate:triethylamine. To the residue containing the product, toluene (200 mL) was evaporated. The yield was 0.15 g.

m/z 460(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ 7.79 (s, 1H), 6.96 (s, 1H), 6.90 (s, 1H), 6.86 (s, 1H), 4.35 (s, 1H), 3.67-3.56 (m, 8H), 3.44 (s, 2H), 2.72 (t, J = 6.7 Hz, 2H), 2.46-2.30 (m, 5H), 1.75-1.66 (m, 2H), 1.58-1.47 ( m, 2H), 1.17 - 1.10 (m, 2H), 1.01 - 0.94 (m, 2H).

e) (R)-7-(2-(3-((4-(2-cyclopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecene) Alkan-9-yl)methyl)-5-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

Trifluoroacetic acid (0.025 mL) was added to (2-cyclopropylthiazol-4-yl)(9-(3-fluoro-5-(2-hydroxyethyl))benzyl at 0 °C in DCM (5 mL) --1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone (Example 57, step d) (0.15 g). The mixture was stirred for 5 min, then Dess-Martin periodinane (0.21 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (5 mL) followed by acetic acid (0.019 mL) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.13 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.031 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 30% acetonitrile). The title compound was obtained as a white solid. The yield was 0.085 g.

m/z 668(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.81 (s, 1H), 7.28-7.16 (m, 3H), 6.93 (d, J = 8.5 Hz, 1H), 6.77 (d, J = 8.2 Hz, 1H), 4.97-4.86 (m, 1H), 4.26 (s, 2H), 3.72-3.58 (m, 6H), 3.31-2.99 (m, 10H), 2.06-1.92 (m, 2H), 1.82 -1.68 (m, 2H), 1.19-0.90 (m, 4H). A proton that is obscured by DMSO and six unobserved exchangeable protons.

Example 58

(R)-7-(2-(3-((4-(2-cyclobutylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 9-yl)methyl)-5-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(cyclobutanecarbamoyl)-3-mercaptopropionic acid ethyl ester

To a suspension of cyclobutanecarboxylic acid (2.57 mL) and 2-amino-3-mercaptopropionic acid ethyl ester hydrochloride (5 g) in DMF (40 mL). The resulting mixture was cooled in an ice-bath, and T.sub.3 solution (1. The resulting mixture was allowed to warm to RT and stirred overnight. The reaction was quenched with saturated sodium bicarbonate solution (50 mL) and partition. The aqueous phase was extracted with DCM (2×100 mL). The combined organic solution was washed with brine (100 mL) dried over sodium sulfate. The white solid formed was purified by chromatography on silica gel eluting with 4:1 to 1:1 isohexane:ethyl acetate gradient. The fractions containing the product are combined and evaporated to give the clear oil of the sub-title compound. The yield was 3.20 g.

m/z 230 (MH)-(APCI) ¹ H NMR (400 MHz, D _{6 -} DMSO) δ 8.05 (d, J = 7.9 Hz, 1H), 4.41-4.32 (m, 1H), 4.14 - 4.05 (m) , 2H), 2.89-2.68 (m, 2H), 2.21-1.70 (m, 7H), 1.18 (t, J = 7.1 Hz, 3H). An unobserved exchangeable proton.

b) Ethyl 2-cyclobutylthiazole-4-carboxylate

Add HCl solution (4M in 1,4-dioxane, 3.46 mL) to ethyl 2-(cyclobutanecarbamoyl)-3-mercaptopropanoate (Example 58, step a) (3.2 g ) in a solution of ethanol (20 mL), and the resulting mixture was stirred overnight. The solvent was evaporated and the residue was azeotroped twice with toluene. The residue was redissolved in acetonitrile (50 mL). EtOAc (EtOAc) The reaction was filtered through a pad of Celite, which was then washed with MeCN (3×100 mL). The combined filtrate and washings were evaporated and the residue was purified by chromatography eluting with EtOAc EtOAc. The fractions containing the product are combined and evaporated to give the clear oil of the sub-title compound. The yield was 0.48 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ8.06 (s, 1H), 4.42 (q, J = 7.1Hz, 2H), 4.00-3.89 (m, 1H), 2.56-2.45 (m, 2H), 2.41- 2.29 (m, 2H), 2.16 - 1.90 (m, 2H), 1.40 (t, J = 7.0 Hz, 3H).

c) 2-cyclobutylthiazole-4-carboxylic acid

To a mixture of THF (8 mL) and water (2 mL) In solution. The resulting suspension was stirred overnight at RT. The reaction was acidified (2M, 5 mL) elut The residue was dissolved between brine (5 mL) and ethyl acetate (20 mL). The layers were separated and the aqueous extracted with EtOAc (2×20 mL). The combined organic solution was dried with sodium sulfate, filtered and evaporated elut The yield was 0.38 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ 12.91 (s, 1H), 8.32 (s, 1H), 3.94-3.83 (m, 1H), 2.46-2.35 (m, 2H), 2.32-2.21 (m, 2H), 2.09-1.97 (m, 1H), 1.94-1.81 (m, 1H).

d) 2-cyclobutylthiazol-4-yl)(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] Undecyl-4-yl)methanone

Adding O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.32g) to 2-(3-(1- Oxa-4,9-diazaspiro[5.5]undecal-9-ylmethyl)-5-fluorophenyl)ethanol (Example 55, step c) (0.2 g), 2-cyclobutane A solution of the thiazole-4-carboxylic acid (Example 58, step c) (0.12 g) and triethylamine (0.36 mL) in DMF (7 mL). The mixture was partitioned between ethyl acetate and brine (100 mL) and layered. The organic phase was washed with brine (2×100 mL) dried over sodium sulfate. The resulting gum was purified by silica gel chromatography eluting with 47.5:47.5:5 isohexane:ethyl acetate:triethylamine to 95:5 ethyl acetate:triethylamine. To the residue containing the product, toluene (200 mL) was added, and the solvent was evaporated to give a subtitle compound. The yield was 0.18 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ 7.91 (s, 1H), 6.95 (s, 1H), 6.89 (s, 1H), 6.86 (s, 1H), 4.36-4.25 (m, 1H ), 3.87 (q, J = 8.3 Hz, 1H), 3.70-3.57 (m, 8H), 3.43 (s, 2H), 2.72 (t, J = 6.7 Hz, 2H), 2.46-2.26 (m, 8H) , 2.11-1.91 (m, 2H), 1.76-1.66 (m, 2H), 1.60-1.51 (m, 2H).

e) (R)-7-(2-(3-((4-(2-cyclobutylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecene) Alkan-9-yl)methyl)-5-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

Add TFA (0.026 mL) to (2-cyclobutylthiazol-4-yl)(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9- Diazaspiro[5.5]undecane-4-yl)methanone (Example 58, step d) (0.16 g) in DCM (5 mL) EtOAc. The mixture was stirred for 5 min, then Dess-Martin periodinane (0.22 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (5 mL) followed by acetic acid (0.019 mL) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.13 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.032 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia solution. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 30% acetonitrile). The title compound was obtained as a white solid. The yield was 0.12 g.

m/z 682 (M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D _{6 -} DMSO, 90 ° C) δ 11.26 (s, 1H), 7.94 (s, 1H), 7.30-7.17 (m, 3H) ), 6.93 (d, J = 8.2 Hz, 1H), 6.78 (d, J = 8.2 Hz, 1H), 4.99 - 4.89 (m, 1H), 4.35 - 4.25 (m, 2H), 3.93 - 3.81 (m, 1H), 3.75-3.62 (m, 6H), 3.32-2.99 (m, 10H), 2.47-2.39 (m, 2H), 2.31-2.20 (m, 2H), 2.10- 1.72 (m, 6H). Five unobserved exchangeable protons.

Example 59

(R)-7-(2-(3-((4-(2-cyclopentylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 9-yl)methyl)-5-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(cyclopentanecarbamido)-3-mercaptopropionic acid ethyl ester

CDI (8.5 g) was added to a solution of cyclopentanecarboxylic acid (5.2 mL) in DMF (40 mL). The resulting mixture was stirred for 1 h and cooled in an ice bath. 2-Amino-3-mercaptopropionic acid ethyl ester hydrochloride (8.88 g) was added followed by triethylamine (10 mL). The resulting mixture was allowed to warm to RT and stirred overnight. The reaction was quenched with saturated sodium bicarbonate solution (50 mL) and partition. The aqueous solution was extracted with DCM (2×100 mL). The combined organic solution was washed with brine (100 mL) dried over sodium sulfate. The solid formed was purified by chromatography on silica gel eluting with 4:1 to 1:1 isohexane:ethyl acetate. The fractions containing the product are combined and evaporated to give the subtitle compound as a white solid. The yield was 7.7 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 8.15 (d, J = 7.7 Hz, 1H), 4.41-4.34 (m, 1H), 4.14 - 4.05 (m, 2H), 2.89-2.61 (m, 3H) ), 1.81-1.45 (m, 9H), 1.18 (t, J = 7.0 Hz, 3H).

b) Ethyl 2-cyclopentyl-4,5-dihydrothiazole-4-carboxylate

p-Toluenesulfonic acid monohydrate (0.78 g) was added to f2-(cyclopentanecarbamimidino)-3-mercaptopropionic acid ethyl ester (Example 59, step a) (5 g) in toluene (40 mL) In the solution. The resulting mixture was heated under reflux for 6 h under Ting-Stark conditions. The reaction was allowed to cool then the toluene solution was washed with sat. The residue was azeotroped with toluene. The white solid formed was purified by chromatography on silica gel eluting with 9:1 isohexane:ethyl acetate. The fractions containing the product are combined and evaporated to give the clear oil of the sub-title compound. The yield is 3.2g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ5.08-4.99 (m, 1H), 4.26 (q, J = 7.2Hz, 2H), 3.56-3.44 (m, 2H), 3.08-2.99 (m, 1H), 2.03-1.93 (m, 2H), 1.83-1.70 (m, 4H), 1.64-1.59 (m, 2H), 1.31 (t, J = 7.2 Hz, 3H).

c) 2-cyclopentylthiazole-4-carboxylic acid ethyl ester

Manganese dioxide (24 g) was added to a solution of 2-cyclopentyl-4,5-dihydrothiazole-4-carboxylic acid ethyl ester (Example 59, step b) (3.2 g) in acetonitrile (100 mL). The resulting mixture was heated under reflux overnight. The reaction was allowed to cool and filtered through a pad of Celite. The filter pad was washed with acetonitrile (2 x 100 mL) and the combined filtrate and washings were evaporated. The residue was purified by silica gel chromatography eluting with 9:1 isohexane:ethyl acetate. The fractions containing the product were combined, evaporated and redissolved in ethanol (20 mL). A slurry of palladium on carbon (5%, 0.66 g) in water (0.5 mL) was added, and the resulting suspension was stirred for 2 h under a 5 bar pressure hydrogen atmosphere. The mixture was filtered through a pad of Celite, which was washed with ethanol (3×50 mL). The combined filtrate and washings were evaporated to give the title compound as a clear oil. The yield was 0.9 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ8.04 (s, 1H), 4.41 (q, J = 7.2Hz, 2H), 3.60-3.48 (m, 1H), 2.29-2.18 (m, 2H), 1.89- 1.67 (m, 6H), 1.40 (t, J = 7.0 Hz, 3H).

d) 2-cyclopentylthiazole-4-carboxylic acid

Add lithium hydroxide monohydrate (0.67 g) to a mixture of 2-cyclopentylthiazole-4-carboxylate (Example 59, step e) (0.9 g) in THF (40 mL) and water (10 mL) In the solution. The resulting mixture was stirred overnight. The reaction was acidified (2M, 10 mL) elut The residue was partitioned between brine (50 mL)EtOAcEtOAc The aqueous phase was extracted with ethyl acetate (2×50 mL). The combined organic solution was dried with sodium sulfate, filtered and evaporated elut The yield was 0.77 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ12.89 (s, 1H), 8.30 (s, 1H), 3.51-3.41 (m, 1H), 2.17-2.07 (m, 2H), 1.82-1.59 (m , 6H).

e) (2-cyclopentylthiazol-4-yl)(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-4-yl)methanone

Adding O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.33g) to 2-(3-(1- Oxa-4,9-diazaspiro[5.5]undecal-9-ylmethyl)-5-fluorophenyl)ethanol (Example 55, step c) (0.21 g), 2-cyclopentane A solution of the thiazole-4-carboxylic acid (Example 59, step d) (0.13 g) and triethylamine (0.38 mL) in DMF (7 mL). The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The resulting gum was purified by silica gel chromatography eluting with 47.5:47.5:5 isohexane:ethyl acetate:triethylamine to 95:5 ethyl acetate:triethylamine. To the residue containing the product, toluene (200 mL) was evaporated. The yield was 0.19 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.89 (s, 1H), 6.95 (s, 1H), 6.88 (s, 1H), 6.86 (s, 1H), 4.30 (t, J = 5.1 Hz, 1H), 3.69-3.61 (m, 8H), 3.43 (s, 2H), 2.76-2.69 (m, 3H), 2.42-2.28 (m, 4H), 2.18-2.06 (m, 2H), 1.86 -1.64 (m, 8H), 1.60-1.49 (m, 2H).

f) (R)-7-(2-(3-((4-(2-)-cyclopentylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecene Alkan-9-yl)methyl)-5-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

Trifluoroacetic acid (0.028 mL) was added to (2-cyclopentylthiazol-4-yl)(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4, 9-Diazaspiro[5.5]undecane-4-yl)methanone (Example 59, step e) (0.18 g) in DCM (5 mL) The mixture was stirred for 5 min, then Dess-Martin periodinane (0.23 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (5 mL) followed by acetic acid (0.021 mL) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.14 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.034 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia solution. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 30% acetonitrile). The title compound was obtained as a white solid. The yield is 0.7g.

m/z 696(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.92 (s, 1H), 7.28-7.15 (m, 3H), 6.93 (d, J = 8.5 Hz, 1H), 6.77 (d, J = 8.2 Hz, 1H), 4.97-4.86 (m, 1H), 4.30-4.20 (m, 2H), 3.74-3.58 (m, 6H), 3.52-3.42 (m, 1H), 3.27 (t, J = 7.9 Hz) , 2H), 3.19-2.96 (m, 8H), 2.18-1.93 (m, 4H), 1.84-1.59 (m, 8H). Six unobserved exchangeable protons.

Example 60

(R)-7-(2-(3-fluoro-5-((4-(4-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5] eleven Cycloalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (9-(3-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(4 -methylthiophen-2-yl)methanone

2,2,2-trifluoro-1-(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] A solution of the carbalide-4-yl)ethanone (Example 41, step d) (0.17 g) in MeOH (3 mL)EtOAc. The mixture was evaporated to dryness under reduced pressure. The residue was dissolved in DMF (6 mL) EtOAc (EtOAc:EtOAc) Stir for 1 hour. The mixture was partitioned between EtOAc and EtOAc. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.120 g.

m/z433(M+H) ⁺ (APCI)

b) (R)-7-(2-(3-Fluoro-5-((4-(4-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

(9-(3-Fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(4-A A solution of the thiophene-2-yl) ketone (Example 60, step a) (0.120 g) in DCM (20 mL) 0.153 g) and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.016 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.109 g) and acetic acid (0.016 mL) in methanol (15 mL). The mixture was cooled in an ice-bath and treated with sodium br The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.075 g.

m/z 641(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.30-7.17 (m, 5H), 6.93 (d, J = 8.2 Hz, 1H), 6.77 (d, J = 8.5 Hz, 1H), 4.94 -4.89(m,1H), 4.27(s,2H),3.73-3.69(m,2H),3.67-3.63(m,2H),3.54(s,2H),3.27(t,J=7.9Hz,2H ), 3.19-2.99 (m, 8H), 2.22 (s, 3H), 2.05-1.97 (m, 2H), 1.82-1.71 (m, 2H). Six unobserved exchangeable protons.

Example 61

(R)-7-(2-(4-chloro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 5-(carboxymethyl)-2-chlorobenzoic acid

Potassium hydroxide (0.969 g) in water (10 mL) was added to a solution of 2-chloro-5-(cyanomethyl)benzoic acid (1.25 g) in ethanol (10 mL). It was heated under reflux for 2.25 hours and then allowed to cool. The mixture was concentrated in vacuo to remove ethanol, then diluted with water and washed twice with ethyl acetate. The organic phase was discarded and the aqueous phase was acidified to pH 1 with concentrated hydrochloric acid and extracted twice with ethyl acetate. The combined extracts were dried over anhydrous MgSO. The yield was 1.38 g. _{^{1 H NMR (400MHz, D 6}} -DMSO) δ 12.93 (br s, 2H), 7.69 (d, J = 2.1Hz, 1H), 7.48 (d, J = 8.3Hz, 1H), 7.42 (dd, J = 8.3, 2.2 Hz, 1H), 3.66 (s, 2H).

b) 2-(4-chloro-3-(hydroxymethyl)phenyl)ethanol

A solution of borane-methylthio complex (2M in THF, 6.50 mL) was added portionwise over 5 min to 5-(carboxymethyl)-2-chlorobenzoic acid (Example 61, step a) (1.37 g) ) in anhydrous THF (20 mL) in a solution at room temperature. The resulting effervescent solution was stirred at room temperature for 1.5 hours and then heated to reflux for 1 hour. The cooled mixture was quenched by the addition of methanol (5 mL) over 5 min. The solution was stirred at room temperature for 2 hours and then purified by flash chromatography eluting EtOAc EtOAc The yield was 0.933 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.36 (d, J = 2.1Hz, 1H), 7.30 (d, J = 7.9Hz, 1H), 7.11 (dd, J = 8.1,2.2Hz, 1H), 4.77 (s, 2H), 3.87 (t, J = 6.5 Hz, 2H), 2.86 (t, J = 6.5 Hz, 2H). Two unobserved exchangeable protons.

c) 2-chloro-5-(2-hydroxyethyl)benzaldehyde

Manganese (IV) (1.00 g) was added to 2-(4-chloro-3-(hydroxymethyl)phenyl)ethanol (Example 61, step b) (0.200 g) in DCM (5 mL) The solution was formed and the resulting suspension was stirred overnight at room temperature. The mixture was then filtered through celite and the residue was washed thoroughly with DCM. The filtrate and washings were concentrated in vacuo to give the subtitle compound as a colourless oil. The yield was 0.197 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ10.47 (s, 1H), 7.80 (d, J = 2.1Hz, 1H), 7.42 (d, J = 2.1Hz, 1H), 7.41 (s, 1H), 3.89 (br t, J = 5.9 Hz, 2H), 2.90 (t, J = 6.4 Hz, 2H), 1.42 (br s, 1H).

d) (9-(2-Chloro-5-(2-hydroxyethyl)benzyl)-4-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -isopropylthiazol-4-yl)methanone

(2-Isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (0.294 g) of a solution in NMP (2 mL). Subsequently a solution of 2-chloro-5-(2-hydroxyethyl)benzaldehyde (Example 61, step c) (0.189 g) in NMP (3 mL) was added and the solution was stirred for one hour and then triethyl hydrazine Treatment with sodium oxyborohydride (0.217 g). The mixture was stirred at room temperature overnight then poured into saturated sodium hydrogen The combined extracts were washed three times with water, once with brine, then dried over anhydrous magnesium sulfate and purified by flash chromatography on ruthenium dioxide, eluting with 1:2:97 triethylamine:methanol:dichloromethane, a colorless gum of the sub-title compound. The yield was 0.238 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.91 (s, 1H), 7.29 (d, J = 1.8 Hz, 1H), 7.25 (d, J = 8.2 Hz, 1H), 7.09 (dd , J=8.2, 2.1 Hz, 1H), 4.29 (t, J=5.1 Hz, 1H), 3.70-3.59 (m, 8H), 3.51 (s, 2H), 3.32 (seven peak, J=6.9 Hz, 1H) ), 2.71 (t, J = 6.8 Hz, 2H), 2.45-2.29 (m, 4H), 1.75-1.66 (m, 2H), 1.60-1.51 (m, 2H), 1.36 (d, J = 6.7 Hz, 6H).

e) 2-(4-Chloro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)methyl)phenyl)acetaldehyde

(9-(2-Chloro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-iso A solution of propyl thiazol-4-yl)methanone (Example 61, step d) (0.230 g) in EtOAc (EtOAc)EtOAc. Dess-Martin periodinane (0.313 g) was added, after which the mixture was taken out from the cooling bath and stirred at room temperature for 40 minutes. The solution was diluted with a saturated aqueous solution of sodium thiosulfate (5 mL), saturated sodium bicarbonate (5 mL) and ethyl acetate (5 mL), and the mixture was stirred vigorously for 10 min. The mixture was then extracted with EtOAc (EtOAc)EtOAc. The yield was 0.274 g.

m/z 476(M+H) ⁺ (APCI)

f) (R)-7-(2-(4-Chloro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-indolylethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

( R )-7-(3-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) A solution of 0.225 g) in MeOH (3 mL) wasEtOAc (EtOAc) Subsequent addition of 2-(4-chloro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 a solution of -methyl)phenyl)acetaldehyde (Example 61, step e) (0.274 g) in MeOH (4 mL), and the mixture was stirred at room temperature for 5 min then cooled in ice-water. Treated with sodium cyanoborohydride (0.051 g). The cooling bath was removed and the mixture was stirred at room temperature for 140 min then treated with more sodium cyanoborohydride (0.055 g). The mixture was stirred overnight. The next morning, the mixture was concentrated in vacuo. The residue was dissolved in a mixture of EtOAc (EtOAc ⁾ (EtOAc ⁾ (EtOAc) The product containing fractions were concentrated in vacuo and co-evaporated three times from acetonitrile to give a colourless residue. The residue was triturated with EtOAc (EtOAc)EtOAc. The yield was 0.062 g.

m/z 686(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.93 (s, 1H), 7.52-7.43 (m, 2H), 7.35-7.26 (m, 1H), 6.93 (d, J = 8.5 Hz, 1H) ), 6.77 (d, J = 8.5 Hz, 1H), 4.89 (dd, J = 7.8, 5.5 Hz, 1H), 3.76-3.60 (m, 6H), 3.36-3.16 (m, 3H), 3.15-2.83 ( m, 8H), 2.02-1.84 (m, 2H), 1.80-1.63 (m, 2H), 1.35 (d, J = 6.9 Hz, 6H). An extremely broad methylene group (two protons) and six unobserved exchangeable protons.

Example 62

(R)-4-hydroxy-7-(1-hydroxy-2-((5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diaza) Spiro[5.5]undecane-9-yl)methyl)naphthalen-1-yl)methylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetaldehyde salt

a) (9-((5-(Bromomethyl)naphthalen-1-yl)methyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)( 2-isopropylthiazol-4-yl)methanone

Add triethylamine (0.185 mL) to (2-isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undec-4-yl)methanone A suspension of trifluoroacetic acid salt (Example 22, step b) (0.225 g) in EtOAc (EtOAc) The residue was dissolved in NMP (5 mL) EtOAc (EtOAc) During this time, a suspension of 1,5-bis(bromomethyl)naphthalene (0.500 g) in acetonitrile (5 mL) and NMP (5 mL) was stirred at room temperature overnight and then concentrated in vacuo to remove acetonitrile to give a solution. The solution of the aforementioned amine was then added dropwise over 35 minutes to complete the addition with more NMP (1 mL). The resulting solution was stirred for 75 minutes, then poured into water and extracted three times with diethyl ether. The combined extracts were washed with water three times and washed with brine, then dried over anhydrous magnesium sulfate and purified by flash chromatography over EtOAc (EtOAc:EtOAc: 50) Dissolution, resulting in a white foam of the sub-title compound. The yield was 0.132 g.

m/z 542/544(M+H) ⁺ (APCI)

b) (R)-4-hydroxy-7-(1-hydroxy-2-((5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxo-4,9-di) Azaspiro[5.5]undecane-9-yl)methyl)naphthalen-1-yl)methylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetic acid salt

Addition of triethylamine (0.16 mL) to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.188 g) in MeOH (5 mL). More triethylamine (0.16 mL) was added dropwise over 30 minutes, together with (9-((5-(bromomethyl)naphthalen-1-yl)methyl)-1-oxa) in NMP (3 mL) -4,9-diazaspiro[5.5]undecyl-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 62, step a) (0.126 g), Multiple NMP (1 mL) was added. The resulting solution was stirred overnight at room temperature. The solution was then filtered, is acidified (0.27 mL) in trifluoroacetic acid and purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile) to borrow. The fractions containing the product were concentrated in vacuo and co-evaporated once from acetonitrile and twice with methanol. The residue was triturated with EtOAc (EtOAc)EtOAc. The yield was 0.056 g.

m/z 688(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.42 (d, J = 8.2 Hz, 1H), 8.31 (brd, J = 8.4 Hz, 1H), 7.93 (s, 1H), 7.87- 7.61 (m, 4H), 6.93 (d, J = 8.5 Hz, 1H), 6.76 (d, J = 8.2 Hz, 1H), 5.00 - 4.94 (m, 1H), 4.75 (dd, J = 23.6, 14.1 Hz) , 2H), 3.80-3.56 (m, 6H), 3.45-2.99 (m, 9H), 2.08-1.87 (m, 2H), 1.81-1.58 (m, 2H), 1.35 (d, J = 6.9 Hz, 6H ). Six unobserved exchangeable protons.

Example 63 (R)-7-(2-(3-Fluoro-5-((4-(5-methylthiophen-2-ylsulfonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(3-Fluoro-5-((4-(5-methylthiophen-2-ylsulfonyl)-1-oxa-4,9-diazaspiro[5.5]undecane -9-yl)methyl)phenyl)ethanol

A solution of 5-methylthiophene-2-sulfonium chloride (0.15 g) in DCM (3 mL) was added dropwise to 2-(3-(1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-ylmethyl)-5-fluorophenyl)ethanol (Example 55, step c) (0.21 g) and N-ethyl- N -isopropylpropan-2-amine (0.14 mL) In a solution of DCM (20 mL). The resulting mixture was stirred for 1 h and the solvent was evaporated. The residue was dissolved in EtOAc (50 mL)EtOAcEtOAc The layers were separated and the aqueous was extracted with ethyl acetate (2×50 mL). The combined organics were washed with brine (50 mL) dried over sodium sulfate. The residue was purified by EtOAc EtOAc (EtOAc:EtOAc:EtOAc: The fractions containing the product are combined and evaporated to give the sub-title compound as a yellow gum. The yield was 0.21 g.

¹ H NMR (400 MHz, D _{6 -} DMSO, 90 ° C) δ 7.46 (d, J = 3.6 Hz, 1H), 7.02 (dd, J = 3.6, 1.0 Hz, 1H), 6.97 (s, 1H), 6.94 ( s, 1H), 6.91 (s, 1H), 4.64 (t, J = 5.1 Hz, 1H), 3.73 - 3.67 (m, 2H), 3.63 - 3.57 (m, 2H), 3.44 (s, 2H), 2.88 -2.83(m,2H),2.74-2.69(m,4H),2.53(s,3H),2.44-2.35(m,2H),2.31-2.22(m,2H),1.80-1.72(m,2H) , 1.60-1.51 (m, 2H)

b) (R)-7-(2-(3-Fluoro-5-((4-(5-methylthiophen-2-ylsulfonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzene well [d]thiazole-2(3H)-one di-trifluoroacetic acid salt

Add TFA (0.033 mL) to 2-(3-fluoro-5-((4-(5-methylthiophen-2-ylsulfonyl)-1-oxa-4,9-diazaspiro[ 5.5]undecane-9-yl)methyl)phenyl)ethanol (Example 63, step a) (0.2 g) in DCM (5 mL) in EtOAc. The mixture was stirred for 5 min, then Dess-Martin periodinane (0.27 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (5 mL) followed by acetic acid (0.024 mL) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.17 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.04 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 30% acetonitrile). The title compound was obtained as a white solid. The yield is 0.7g.

m/z 677(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.43 (d, J = 3.7 Hz, 1H), 7.30-7.16 (m, 3H), 7.00-6.96 (m, 1H), 6.93 (d, J=8.5 Hz, 1H), 6.78 (d, J=8.3 Hz, 1H), 4.92 (dd, J=7.9, 5.4 Hz, 1H), 4.29 (s, 2H), 3.79-3.71 (m, 2H), 3.32-3.23 (m, 2H), 3.17-2.93 (m, 10H), 2.85 (s, 2H), 2.53 (s, 3H), 2.13-2.02 (m, 2H), 1.90-1.74 (m, 2H). Six unobserved exchangeable protons.

Example 64

(R)-4-hydroxy-7-(1-hydroxy-2-(2-(3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxo-4,9-di) Azaspiro[5.5]undecane-9-yl)methyl)phenoxy)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (9-(3-(2,2-diethoxyethoxy)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl) (2-isopropylthiazol-4-yl)methanone

Methanesulfonium chloride (0.110 mL) was added dropwise to (3-(2,2-diethoxyethoxy)phenyl)methanol (Example 38, Step a) (0.307 g) and triethylamine ( 0.196 mL) in a stirred solution of 0 ° C in DCM (30 mL). The resulting mixture was stirred at 20 ° C for 1 hour. The mixture was washed with water and the org. The residue was dissolved in acetonitrile (30 mL) eluted with triethylamine (1 mL) then (2- isopropyl thiazol-4-yl) Treatment with undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (0.400 g). The mixture was stirred at 20 ° C for 2 hours. The solvent was removed under reduced pressure and the residue was partitioned between ethyl acetate and brine. The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.360 g.

m/z 532(M+H) ⁺ (APCI)

b) 2-(3-((4-(2-Isopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)) Phenoxy)acetaldehyde

(9-(3-(2,2-Diethoxyethoxy)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -Isopropylthiazole-4-yl)methanone (Example 64, step a) (0.36 g) in EtOAc (20 mL) EtOAc (EtOAc) Heat for 18 hours. The solvent was evaporated under reduced pressure. The yield was 0.31 g. Use directly.

c) (R)-4-hydroxy-7-(1-hydroxy-2-(2-(3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxo-4,9) -diazaspiro[5.5]undecane-9-yl)methyl)phenoxy)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetic acid salt

2-(3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl) a solution of phenoxy)acetaldehyde (Example 64, step b) (0.31 g) in methanol (10 mL) with ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzene And [d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.267 g), followed by acetic acid (0.039 mL), and the mixture was cooled in ice. Sodium cyanoborohydride (0.085 g) was added, and the mixture was stirred at room temperature for 4 hr. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.2 g.

m/z 668(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.27 (s, 1H), 7.94 (s, 1H), 7.39 (t, J = 8.0Hz, 1H), 7.16-7.13 (m, 2H) , 7.09-7.05 (m, 1H), 6.94 (d, J = 8.5 Hz, 1H), 6.78 (d, J = 8.2 Hz, 1H), 4.98 - 4.93 (m, 1H), 4.36-4.28 (m, 4H) ), 3.70 (s, 4H), 3.65 (s, 2H), 3.47-3.42 (m, 2H), 3.34 - 3.26 (m, 1H), 3.23 - 3.05 (m, 6H), 2.09 - 1.98 (m, 2H) ), 1.84-1.71 (m, 2H), 1.35 (d, J = 7.1 Hz, 6H). Five unobserved exchangeable protons.

Example 65

(R)-4-mercapto-7-(1-hydroxy-2-(2-(3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxo-4,9-) Diazaspiro[5.5]undecane-9-yl)methyl)phenylthio)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetic acid salt

a) 3-(2-( third -butyldimethylmethylalkyloxy)ethylthio)benzoic acid

(2-Bromoethoxy)(t-butyl)dimethyl decane (1.48 mL) was added dropwise to 3-mercaptobenzoic acid (1.07 g) and potassium carbonate (1.91 g) in DMF (15 mL) In suspension. The resulting suspension was stirred for 2 h. The reaction was cautiously acidified (2M, 10 mL) eluting EtOAc (EtOAc) The resulting aqueous solution was extracted with ethyl acetate (3×100 mL). The combined organics were washed with EtOAc (EtOAc)EtOAc. The yield was 2.8 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 13.11 (s, 1H), 7.89 (t,J = 1.7 Hz, 1H), 7.79-7.74 (m, 1H), 7.66-7.61 (m, 1H), 7.47 (t, J = 7.7 Hz, 1H), 3.82 (t, J = 6.4 Hz, 2H), 3.19 (t, J = 6.4 Hz, 2H), 0.87 (s, 9H), 0.05 (s, 6H).

b) 2-(3-(hydroxymethyl)phenylthio)ethanol

Borane dimethylsulfide complex compound (in THF 2M, 17.3mL) was added dropwise to 3- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethylthio) benzoic acid ( Example 65, step a) (2.16 g) in EtOAc (EtOAc) The reaction was allowed to warm to RT then heated at reflux for 2 h. The reaction was cooled in an ice-bath and aqueous HCl (2M, 50 mL). The resulting mixture was stirred overnight. The reaction was concentrated to one half of its original volume and the resulting aqueous solution was extracted with DCM (3×100 mL). The combined organics were washed with brine (100 mL) dried over sodium sulfate. The residue was purified by EtOAc (EtOAc) elute The fractions containing the product are combined and evaporated to give the clear oil of the sub-title compound. The yield was 0.77 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ 7.29-7.23 (m, 2H), 7.21-7.17 (m, 1H), 7.13-7.09 (m, 1H), 5.20 (t, J = 5.8Hz, 1H) , 4.92 (t, J = 5.6 Hz, 1H), 4.47 (d, J = 5.9 Hz, 2H), 3.59 - 3.53 (m, 2H), 3.02 (t, J = 6.9 Hz, 2H).

c) 3-(2-hydroxyethylthio)benzaldehyde

Manganese dioxide (1.36 g) was added to a solution of 2-(3-(hydroxymethyl)phenylthio)ethanol (Example 65, step b) (0.29 g) in DCM (10 mL). The resulting mixture was heated under reflux for 4 h, cooled and filtered over EtOAc. The filter pad was washed with DCM (3 x 20 mL). The filtrate and washings were combined and evaporated to give the sub-title compound as a yellow gum. The yield was 0.2 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 9.99 (s, 1H), 7.84 (s, 1H), 7.71-7.64 (m, 2H), 7.54 (t, J = 7.7 Hz, 1H), 4.99 ( t, J = 5.6 Hz, 1H), 3.64 - 3.57 (m, 2H), 3.13 (t, J = 6.7 Hz, 2H).

d) (9-(3-(2-hydroxyethylthio)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-iso Propylthiazole-4-yl)methanone

3-(2-Hydroxyethylthio)benzaldehyde (Example 65, step c) (0.16 g) was added to (2-isopropylthiazol-4-yl)(1-oxa-4,9- Diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (0.25 g) and acetic acid (0.046 mL) in N-methyl-2-pyrrolidine In a solution of the ketone (5 mL), the resulting mixture was stirred for 1 h and cooled in ice. Sodium triethoxysulfonate (0.26 g) was then added and the mixture was warmed to RT and stirred overnight. The reaction was poured into a mixture of sodium bicarbonate solution (20 mL) and water (EtOAc). The aqueous phase was extracted with diethyl ether (3×100 mL). The combined organic solution was washed with brine (100 mL) dried over sodium sulfate. The crude product was purified by silica gel chromatography eluting with 95:5 ethyl acetate: triethylamine. The fractions containing the product are combined and evaporated to give the sub-title compound as a yellow oil. The yield was 0.22 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.94-7.83 (m, 1H), 7.28-7.18 (m, 3H), 7.12-7.05 (m, 1H), 4.61-4.53 (m, 1H) ), 3.70-3.54 (m, 8H), 3.47-3.39 (m, 2H), 3.37-3.27 (m, 1H), 3.06-2.96 (m, 2H), 2.42-2.25 (m, 4H), 1.75-1.65 (m, 2H), 1.59-1.50 (m, 2H), 1.41-1.32 (m, 6H).

e) (R)-4-hydroxy-7-(1-hydroxy-2-(2-(3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxo-4,9) -diazaspiro[5.5]undecal-9-yl)methyl)phenylthio)ethylamino)ethyl)benzene well [d]thiazole-2(3H)-one di-trifluoro Acetate

Trifluoroacetic acid (0.033 mL) was added to (9-(3-(2-hydroxyethylthio)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 4-Base)(2-Isopropylthiazol-4-yl)methanone (Example 65, step d) (0.22 g) in DCM (5 mL) The mixture was stirred for 5 min, then Dess-Martin periodinane (0.29 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (5 mL) followed by acetic acid (0.026 mL) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.15 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.044 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 30% acetonitrile). The title compound was obtained as a white solid. The yield was 0.54 g.

m/z 684(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.94 (s, 1H), 7.54 - 7.34 (m, 4H), 6.91 (d, J = 8.2 Hz, 1H), 6.76 (d, J = 8.2 Hz, 1H), 4.88 (dd, J=8.7, 4.6 Hz, 1H), 4.24 (s, 2H), 3.74-3.61 (m, 6H), 3.38-2.95 (m, 11H), 2.08-1.94 (m) , 2H), 1.82-1.68 (m, 2H), 1.35 (d, J = 6.9 Hz, 6H). Six unobserved exchangeable protons.

Example 66

(R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(4-isopropylthiazol-2-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trioxyacetate

a) 4-(4-isopropylthiazol-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester

Add HATU (0.924g) to 4-isopropylthiazole-2-carboxylic acid (0.32g) and 1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid at a time. Butyl ester (Example 12, step b) (0.547 g) and triethylamine (0.781 mL) in DMF (12 mL). The reaction mixture was stirred at room temperature for 2 hours. The mixture was partitioned between EtOAc and EtOAc. The crude product was purified by flash chromatography on silica gel eluting with 30% ethyl acetate in isohexane. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.54 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ 7.50 (s, 1H), 3.77-3.72 (m, 2H), 3.52-3.45 (m, 2H), 3.21-3.13 (m, 2H), 3.12 -3.04 (m, 1H), 3.00 (s, 4H), 1.74-1.67 (m, 2H), 1.52-1.43 (m, 2H), 1.39 (s, 9H), 1.26 (d, J = 6.8 Hz, 6H) ).

b (4-isopropylthiazol-2-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone

3-(4-Isopropylthiazole-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester (Example 66, step a The solution (0.54 g) in DCM (20 mL) wasEtOAc. Toluene (40 mL) was added and the solvent was removed under reduced pressure. The residue was azeotroped twice with acetonitrile to give the subtitle compound. The yield was 0.56 g.

m/z 310(M+H) ⁺ (APCI)

c) (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(4-isopropyl Thiazol-2-yl)methanone

(4-Isopropylthiazol-2-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone (Example 66, Step b) (0.28 g) and 2-(3-(bromomethyl)phenyl)ethanol (Example 6, step a) (0.171 g) in acetonitrile (20 mL) EtOAc (EtOAc) Stir at 20 ° C for 2 hours. The solvent was evaporated under reduced pressure. The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The crude product was purified by flash chromatography using EtOAc (EtOAc): The yield is 0.25 g.

m/z 444(M+H) ⁺ (APCI)

d) (R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(4-isopropylthiazole-2-carbonyl)-1-oxa-4,9-diaza) Hetero[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(4-isopropylthiazole- A solution of 2-yl)methanone (Example 66, step c) (0.25 g) in EtOAc (EtOAc) The resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.032 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.222 g) and acetic acid (0.032 mL) in MeOH (20 mL). The mixture was cooled in an ice-bath and treated with sodium <RTI ID=0.0> The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired product were evaporated to dryness to give the title compound. The yield was 0.18 g.

m/z 652(M+H) ⁺ (APCI)

_{^{1 H NMR (300MHz, D 6}} -DMSO, 90 ℃) δ11.35 (s, 1H), 7.53 (s, 1H), 7.43-7.32 (m, 4H), 6.93 (d, J = 8.3Hz, 1H) , 6.78 (d, J = 8.5 Hz, 1H), 4.97-4.89 (m, 1H), 4.29 (s, 2H), 3.76 (s, 2H), 4.17-3.30 (m, 4H), 3.28-3.15 (m , 4H), 3.14 - 2.98 (m, 7H), 2.10 - 1.97 (m, 2H), 1.87-1.70 (m, 2H), 1.26 (d, J = 7.1 Hz, 6H). Five unobserved exchangeable protons.

Example 67

(R)-7-(2-(2-Fluoro-3-((4-(2-(2,2,2-trifluoroethyl)thiazole-4-carbonyl)-1-oxa-4,9 -diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one Di-trifluoroacetate

a) 3,3,3-trifluoropropane acid amine

A solution of 3,3,3-trifluoropropionic acid (4 g) in diethyl ether (150 mL) was taken &lt The mixture was heated under nitrogen for 2.5 hours under reflux. The reaction mixture was cooled to room temperature. Additional diethyl ether (100 mL) was added and the mixture was cooled in ice. Ammonia was bubbled through the mixture for 30 minutes. The solvent was removed under reduced pressure and the residue was crystallised eluted eluted eluted The aqueous layer was re-extracted twice with ethyl acetate. EtOAc (EtOAc) The yield was 3.9 g. Use directly.

b) 3,3,3-trifluoropropanethioguanamine

Phosphorus pentasulfide (1.603 g) was added in one portion to a solution of 3,3,3-trifluoropropanoguanamine (Example 67, step a) (3.9 g) in MTBE (150 mL). The reaction mixture was stirred at 20 ° C for 20 hours. The mixture was filtered and the solvent was evaporated <RTI ID=0.0> The yield is 4g. The product was used directly in the next step.

c) ethyl 2-(2,2,2-trifluoroethyl)thiazole-4-carboxylate

a solution of 3,3,3-trifluoropropanethioguanamine (Example 67, step b) (4 g) and ethyl 3-bromo-2-oxopropionate (5.45 g) in THF (120 mL) Heat under reflux for 2 hours. The solvent was removed under reduced pressure and the residue was partitioned between ethyl acetate and saturated sodium hydrogen carbonate. The organic layer was dried with sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 1.9 g.

m/z 240(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, CDC1 3}} ) δ 8.23 (s, 1H), 4.45 (q, J = 7.1Hz, 2H), 3.94 (q, J = 10.2Hz, 2H), 1.42 (t, J = 7.0Hz, 3H).

d ) 2-(2,2,2-trifluoroethyl)thiazole-4-carboxylic acid

a mixture of ethyl 2-(2,2,2-trifluoroethyl)thiazole-4-carboxylate (Example 67, step c) (0.3 g) in EtOAc (EtOAc) Heat under nitrogen for 5 hours. The solvent was evaporated under a nitrogen stream and the residue was partitioned between ethyl acetate (40mL) and brine (3mL). The organic layer was dried with sodium sulfate, filtered and evaporatedEtOAc The yield was 0.23 g.

m/z 210 (M-H) (APCI)

¹ H NMR (300 MHz, CDCl ₃ ) δ 8.36 (s, 1H), 3.95 (q, J = 10.0 Hz, 2H). An unobserved exchangeable proton.

e) 2,2,2-trifluoro-1-(9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] Undecyl-4-yl)ethanone

2-Fluoro-3-(2-hydroxyethyl)benzaldehyde (Example 48, step a) (1.05 g) was added to 2,2,2-trifluoro-1-(1-oxa-4,9 -Diazaspiro[5.5]undecane-4-yl)ethanone trifluoroacetate (Example 12, step d) (2.08 g) and acetic acid (0.33 mL) in N-methyl-2-pyrrole In a solution of ketone (10 mL). The resulting mixture was stirred for 15 min and then cooled in an ice bath. Sodium triethoxy borohydride (1.81 g) was then added and the mixture was stirred overnight. The reaction was poured into a mixture of saturated sodium bicarbonate (20 mL) and water (100 mL). The aqueous solution was extracted with ethyl acetate (3×100 mL). The combined organics were washed with water (50 mL) brine brine The residue was purified by EtOAc (EtOAc) elute The fractions containing the product are combined and evaporated to give the clear oil of the sub-title compound. The yield was 1.9 g. use immediately.

f) 2-(3-(1-oxa-4,9-diazaspiro[5.5]undecane-9-ylmethyl)-2-fluorophenyl)ethanol

Add 880' aqueous ammonia solution (5 mL) to 2,2,2-trifluoro-1-(9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9 -diazaspiro[5.5]undecane-4-yl)ethanone (Example 67, step e) (1.9 g) in MeOH (25 mL). The resulting mixture was stirred for 90 min and the solvent was evaporated. The residue was purified by silica gel chromatography eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM: methanol: &lt; The fractions containing the product are combined and evaporated to yield a gum which solidifies upon standing. The white solid was triturated with EtOAc (EtOAc) elute The yield is 1g.

m/z 309(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO) δ 7.24 - 7.15 (m, 2H), 7.06 (t,J = 7.6 Hz, 1H), 4.69 (t, J = 5.3 Hz, 1H), 3.64 - 3.54 ( m, 2H), 3.53-3.43 (m, 4H), 2.74 (t, J = 7.0 Hz, 2H), 2.60 (t, J = 4.9 Hz, 2H), 2.45 - 2.35 (m, 2H), 2.34 - 2.24 (m, 2H), 1.83-1.74 (m, 2H), 1.47-1.36 (m, 2H). Two protons obscured by solvent peaks and one unobserved exchangeable proton.

g) (9-(2-Fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -(2,2,2-triazoethyl)thiazol-4-yl)methanone

Add HATU (0.181 g) once to 2-(2,2,2-trifluoroethyl)thiazole-4-carboxylic acid (Example 67, step d) (0.077 g) and 2-(3-(1-oxygen) Hetero-4,9-diazaspiro[5.5]undecal-9-ylmethyl)-2-fluorophenyl)ethanol (Example 67, step f) (0.113 g) and triethylamine (0.153) mL) in a stirred solution of 0 ° C in DMF (5 mL). The reaction mixture was stirred at room temperature for 2 hours. The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The yield was 0.180 g.

m/z 502(M+H) ⁺ (APCI)

h) (R)-7-(2-(2-Fluoro-3-((4-(2-(2,2,2-trifluoroethyl)thiazole-4-carbonyl)-1-oxa-4 ,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H) -keto-difluoroacetate

(9-(2-Fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-( A solution of 2,2,2-trifluoroethyl)thiazol-4-yl)methanone (Example 67, step g) (0.18 g) inEtOAc (EtOAc) The mixture was treated with Dess-Martin periodinane (0.198 g) and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.021 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.141 g) and acetic acid (0.021 mL) in methanol (15.0 mL). The mixture was cooled in an ice-bath and treated with sodium <RTI ID=0.0> The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.091 g.

m/z 710(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.15 (s, 1H), 7.52-7.40 (m, 2H), 7.25 (t, J = 7.6 Hz, 1H), 6.94 (d, J = 8.2 Hz, 1H), 6.77 (d, J = 8.2 Hz, 1H), 4.94 - 4.89 (m, 1H), 4.29 (s, 2H), 4.20 (q, J = 11.0 Hz, 2H), 3.74 - 3.61 ( m, 6H), 3.28-3.02 (m, 10H), 2.06-1.96 (m, 2H), 1.85-1.72 (m, 2H). Six unobserved exchangeable protons.

Example 68

(R)-7-(2-(3-((4-(benzo[b]thiophen-5-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 9-yl)methyl)-2-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) benzo[b]thiophen-5-yl(9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkan-4-yl)methanone

Add HATU (0.181 g) to benzo[b]thiophene-5-carboxylic acid (0.065 g) and 2-(3-(1-oxa-4,9-diazaspiro[5.5]undecane in one portion. -9-Methylmethyl)-2-fluorophenyl)ethanol (Example 67, step f) (0.113 g) and triethylamine (0.153 mL). The reaction mixture was stirred at room temperature for 2 hours. The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The yield was 0.160 g.

m/z 469(M+H) ⁺ (APCI)

b) (R)-7-(2-(3-((4-(benzo[b]thiophene-5-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undec carbon Alkan-9-yl)methyl)-2-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

Benzo[b]thiophen-5-yl(9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undec carbon Alkyl-4-yl)methanone (Example 68, step a) (0.160 g) EtOAc (EtOAc m. g) and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.02 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.135 g) and acetic acid (0.02 mL) in methanol (15 mL). The mixture was cooled in an ice-bath and treated with sodium br The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.116 g.

m/z 677(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D _{6 -} DMSO, 90 ° C) δ 11.28 (s, 1H), 8.04 (d, J = 8.2 Hz, 1H), 7.91 (s, 1H), 7.80 (d, J = 5.4 Hz, 1H), 7.52-7.42 (m, 3H), 7.38-7.35 (m, 1H), 7.25 (t, J = 7.7 Hz, 1H), 6.94 (d, J = 8.5 Hz, 1H), 6.78 (t, J) =4.1 Hz, 1H), 4.95-4.90 (m, 1H), 4.35 (s, 2H), 3.69 (t, J = 5.0 Hz, 2H), 3.53-3.44 (m, 4H), 3.28-3.19 (m, 4H), 3.16-3.04 (m, 6H), 2.11-2.03 (m, 2H), 1.80-1.68 (m, 2H). Five unobserved exchangeable protons.

Example 69

(R)-7-(2-(2-fluoro-3-((4-(2-isopropyl-5-methylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoro Acetate

a) methyl 2-isopropyl-5-methylthiazole-4-carboxylate

A mixture of methyl 3-bromo-2-oxobutanoate (4.6 g) and 2-methylpropanesulfonamide (2.5 g) in THF (100 mL) The solvent was evaporated under reduced pressure. The residue was partitioned between EtOAc EtOAc m. The residue was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 1.7 g.

m/z 200(M+H) ⁺ (APCI)

b) 2-isopropyl-5-methylthiazole-4-carboxylic acid

A solution of lithium hydroxide monohydrate (0.2 g) in water (3 mL) was added to methyl 2-isopropyl-5-methylthiazole-4-carboxylate (Example 69, step a) (0.5 g) In a solution of methanol (7 mL), and the mixture was stirred at 20 ° C for 3 hr. Methanol was removed under reduced pressure and the remaining aqueous solution was washed with ethyl acetate. The aqueous layer was acidified by dropwise addition of concentrated aqueous hydrochloric acid and extracted with ethyl acetate. The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The formed gel crystallizes upon standing. Wet milling with a mixture of isohexane (4 mL) and diethyl ether (1 mL) gave subtitle compound. The yield was 0.160 g.

¹ H NMR (400 MHz, CDC1 ₃ ) δ 3.29 - 3.18 (m, 1H), 2.78 (s, 3H), 1.38 (d, J = 7.1 Hz, 6H). An unobserved exchangeable proton.

c) (9-(2-Fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -isopropyl-5-methylthiazol-4-yl)methanone

HATU (0.175 g) was added in one portion to 2-isopropyl-5-methylthiazole-4-carboxylic acid (Example 69, step b) (0.066 mg) and 2-(3-(1-oxo-4). 9-diazaspiro[5.5]undecal-9-ylmethyl)-2-fluorophenyl)ethanol (Example 67, step f) (0.109 g) and triethylamine (0.148 mL) in DMF (5 mL) in a stirred solution at 0 °C. The reaction mixture was stirred at room temperature for 2 hours. The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The yield was 0.155 mg.

m/z 476(M+H) ⁺ (APCI)

d) (R)-7-(2-(2-fluoro-3-((4-(2-isopropyl-5-methylthiazole-4-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di- Trifluoroacetate

(9-(2-Fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-iso A solution of propyl-5-methylthiazol-4-yl)methanone (Example 69, step c) (0.155 g) in DCM (15 mL Martin was treated with iodosane (0.180 g) and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.019 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.128 g) and acetic acid (0.019 mL) in methanol (15.00 mL). The mixture was cooled in an ice-bath and treated with sodium br The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.12 g.

m/z 684(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.28 (s, 1H), 7.50-7.39 (m, 2H), 7.28-7.22 (m, 1H), 6.93 (d, J = 24.1Hz, 1H), 6.77 (d, J = 26.1 Hz, 1H), 4.93-4.88 (m, 1H), 4.26 (s, 2H), 3.67 (s, 2H), 3.58-3.43 (m, 4H), 3.26-3.00 (m, 11H), 2.43 (s, 3H), 2.03-1.94 (m, 2H), 1.81-1.65 (m, 2H), 1.30 (d, J = 6.8 Hz, 6H). Five unobserved exchangeable protons.

Example 70

( R )-7-(2-(2-Fluoro-3-((4-(5-isopropylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (9-(2-Fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(5 -isopropylthiophen-3-yl)methanone

Add HATU (0.183 g) to 5-isopropylthiophene-3-carboxylic acid (0.063 g) and 2-(3-(1-oxa-4,9-diazaspiro[5.5]undecane in one portion. A stirred solution of -9-ylmethyl)-2-fluorophenyl)ethanol (Example 67, step f) (0.114 g) and triethylamine (0.155 mL) in DMF (5 mL) The reaction mixture was stirred at room temperature for 2 hours. The mixture was partitioned between EtOAc and EtOAc. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.100 g.

m/z 461(M+H) ⁺ (APCI)

b) (R)-7-(2-(2-Fluoro-3-((4-(5-isopropylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

(9-(2-Fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(5-iso A solution of propylthiophen-3-yl)methanone (Example 70, step a) (0.100 g) in DCM (15 mL) (0.120 g) and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.012 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.086 g) and acetic acid (0.012 mL) in methanol (15.00 mL). The mixture was cooled in an ice-bath and dried (EtOAc) The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.052 mg.

m/z 669(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.51 - 7.46 (m, 2H), 7.43 (t, J = 7.5 Hz, 1H), 7.25 (t, J = 7.7 Hz, 1H), 6.95 -6.89 (m, 2H), 6.77 (d, J = 8.2 Hz, 1H), 4.95 - 4.90 (m, 1H), 4.30 (s, 2H), 3.69 - 3.65 (m, 2H), 3.55 - 3.51 (m , 2H), 3.45 (s, 2H), 3.24 (t, J = 8.1 Hz, 2H), 3.20-3.03 (m, 9H), 2.05-1.97 (m, 2H), 1.79-1.69 (m, 2H), 1.28 (d, J = 6.8 Hz, 6H). Six unobserved exchangeable protons.

Example 71 (R)-7-(2-(2-Fluoro-3-((4-(2-(pentan-3-yl)thiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoro Acetate

a) 2-ethylbutane decylamine

2-Ethyl butane ruthenium chloride (5 g) was added cautiously to ice-cold 35% aqueous ammonia (50 mL) and the resulting mixture was stirred for 1 h. The reaction mixture was extracted with DCM (3×100 mL). The combined organics were washed with EtOAc EtOAc m. The yield was 3.4 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ7.23 (s, 1H), 6.71 (s, 1H), 1.98-1.88 (m, 1H), 1.50-1.27 (m, 4H), 0.81 (t, J =7.4HZ, 6H).

b) 2-ethylbutane sulfate

Phosphorus pentasulfide (1.54 g) was added to a solution of 2-ethylbutane decylamine (Example 71, step a) (3.4 g) in MTBE (300 mL). The reaction was filtered through celite and the pad was washed with EtOAc (EtOAc). The combined filtrate and washings were evaporated to give the subtitle compound as a yellow oil. The yield was 3.8 g. Use directly.

c) ethyl 2-(pentan-3-yl)thiazole-4-carboxylate

Ethyl 3-bromo-2-oxopropionate (2.5 mL) was added dropwise to 2-ethylbutanethioguanamine (Example 71, step b) (3.8 g) in ethanol (100 mL) The solution was formed and the mixture formed was heated under reflux overnight. The solvent was evaporated and the residue was crystalljjjjjjjjjjjj The layers were separated and the aqueous extracted with ethyl acetate (2×100 mL). The combined organic solution was washed with brine (100 mL) dried over sodium sulfate. The residue was purified by silica gel chromatography eluting with 20:1 isohexane:ethyl acetate. The fractions containing the product are combined and evaporated to give the sub-title compound as a yellow oil. The yield was 2.8 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ8.42 (s, 1H), 4.29 (q, J = 7.1Hz, 2H), 2.99-2.89 (m, 1H), 1.82-1.58 (m, 4H), 1.30 (t, J = 7.0 Hz, 3H), 0.81 (t, J = 7.4 Hz, 6H).

d) 2-(pentan-3-yl)thiazole-4-carboxylic acid

Lithium hydroxide monohydrate (2.07 g) was added to ethyl 2-(pentan-3-yl)thiazole-4-carboxylate (Example 71, step c) (2.8 g) in THF (80 mL) In a solution of a mixture of 20 mL). The resulting mixture was stirred overnight. The reaction was acidified with concentrated hydrochloric acid (6 mL) and evaporated. The aqueous mixture formed was saturated with sodium chloride and extracted with ethyl acetate (3×100 mL). The combined organic solution was dried with sodium sulfate, filtered and evaporated elut The yield was 2.3 g.

_{^{1 H NMR (300MHz, D 6}} -DMSO) δ12.91 (s, 1H), 8.34 (s, 1H), 2.98-2.86 (m, 1H), 1.84-1.56 (m, 4H), 0.81 (t, J = 7.3 Hz, 6H).

e) (9-(2-Fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -(pentan-3-yl)thiazol-4-yl)methanone

Adding O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.2g) to 2-(3-(1- Oxa-4,9-diazaspiro[5.5]undecal-9-ylmethyl)-2-fluorophenyl)ethanol (Example 67, step f) (0.13 g), 2-(pent Alkyl-3-yl)thiazole-4-carboxylic acid (Example 71, step d) (0.081 g) and triethylamine (0.23 mL) in DMF (7 mL) in EtOAc. The resulting yellow solution was allowed to warm to RT and stirred for 2 h. The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The resulting gum was purified by silica gel chromatography eluting with 47.5:47.5:5 isohexane:ethyl acetate:triethylamine to 95:5 ethyl acetate:triethylamine. To the residue containing the product, toluene (200 mL) was evaporated. The yield is 0.25 g.

m/z 490(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.93 (s, 1H), 7.22 - 7.14 (m, 2H), 7.03 (t, J = 7.6 Hz, 1H), 4.39-4.32 (m, 1H), 3.69-3.58 (m, 6H), 3.52-3.46 (m, 2H), 2.97-2.90 (m, 1H), 2.75 (t, J = 7.3 Hz, 2H), 2.70 (s, 2H), 2.46 - 2.29 (m, 4H), 1.82-1.66 (m, 6H), 1.58-1.47 (m, 2H), 0.85 (t, J = 7.3 Hz, 6H).

f) (R)-7-(2-(2-Fluoro-3-((4-(2-(pentan-3-yl)thiazole-4-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di- Trifluoroacetate

Adding trifluoroacetic acid (0.031 mL) to (9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undec carbon Alkyl-4-yl)(2-(pentan-3-yl)thiazol-4-yl)methanone (Example 71, step e) (0.2 g) in DCM (5 mL) The mixture was stirred for 5 min, then Dess-Martin periodinane (0.25 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (5mL), followed by addition of acetic acid (0.023 mL) and (R) -7- (2- amino-1-hydroxyethyl) -4-hydroxybenzo [d] thiazol -2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.13 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.038 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 35% acetonitrile). The title compound was obtained as a white solid. The yield was 0.17 g.

m/z 698(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D _{6 -} DMSO, 90 ° C) δ 11.26 (s, 1H), 8.81 - 7.83 (m, 1H), 7.56-7.38 (m, 2H), 7.30-7.19 (m, 1H), 7.01 -6.89(m,1H),6.83-6.71(m,1H),4.99-4.88(m,1H), 4.38-4.23(m,2H), 3.78-3.57(m,6H),3.33-2.86(m, 11H), 2.10 - 1.97 (m, 2H), 1.87-1.61 (m, 6H), 0.91 - 0.76 (m, 6H). Five unobserved exchangeable protons.

Example 72

(R)-7-(2-(2-fluoro-3-((4-(2-isobutylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 3-methylbutane amide

3-Methylbutane ruthenium chloride (10 mL) was carefully added dropwise to ice cold 35% aqueous ammonia (50 mL) and the resulting mixture was stirred for 1 h. The reaction mixture was extracted with DCM (3×100 mL). The combined organics were washed with EtOAc (EtOAc)EtOAc. The yield was 5.6 g.

¹ H NMR (300 MHz, D _{6 -} DMSO) δ 7.20 (s, 1H), 6.67 (s, 1H), 1.99-1.88 (m, 3H), 0.87 (d, J = 6.4 Hz, 6H).

b) 3-methylbutane sulfate

Phosphorus pentasulfide (2.9 g) was added to a suspension of 3-methylbutaneamine (Example 72, step a) (5.6 g) in EtOAc (300 mL). The reaction was filtered through celite and the pad was washed with EtOAc (EtOAc). The combined filtrate and washings were evaporated to give the subtitle compound as a yellow oil. The yield was 5.6 g.

_{^{1 H NMR (300MHz, D 6}} -DMSO) δ9.32 (s, 1H), 9.10 (s, 1H), 2.33 (d, J = 7.3Hz, 2H), 2.22-2.07 (m, 1H), 0.88 ( d, J = 6.4 Hz, 6H).

c) 2-isobutylthiazole-4-carboxylic acid ethyl ester

To a solution of 3-methylbutanethioguanamine (Example 72, step b) (5.6 g) in ethanol (100 mL), ethyl 3-bromo-2-oxopropionate (6.7 mL). The resulting mixture was stirred at RT overnight and then heated at reflux for 5 h. The solvent was evaporated and the residue was crystalljjjjjjjjjjjjj The layers were separated and brine (100 mL)EtOAc. The residue was purified by silica gel chromatography eluting with 20:1 to 10:1 isohexane:EtOAc. The fractions containing the product are combined and evaporated to give the sub-title compound as a yellow oil. The yield was 5.2 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ8.39 (s, 1H), 4.29 (q, J = 7.1Hz, 2H), 2.88 (d, J = 7.2Hz, 2H), 2.10-1.97 (m, 1H), 1.30 (t, J = 7.2 Hz, 3H), 0.93 (d, J = 6.7 Hz, 6H).

d) 2-isobutylthiazole-4-carboxylic acid

Add lithium hydroxide monohydrate (4.1 g) to a mixture of 2-isobutylthiazole-4-carboxylate (Example 72, step c) (5.2 g) in THF (80 mL) and water (20 mL) In the solution. The resulting mixture was stirred overnight. The reaction was carefully acidified (10 mL) with concentrated hydrochloric acid and evaporated. The resulting aqueous mixture was poured into brine (50 mL) andEtOAcEtOAc The combined organics were washed with EtOAc (EtOAc)EtOAc. The yield was 3.8 g.

_{^{1 H NMR (300MHz, D 6}} -DMSO) δ12.90 (s, 1H), 8.32 (s, 1H), 2.87 (d, J = 7.1Hz, 2H), 2.11-1.96 (m, 1H), 0.94 ( d, J = 6.7 Hz, 6H).

e) (9-(2-Fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -isobutylthiazole-4-yl)methanone

Adding O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.2g) to 2-(3-(1- Oxa-4,9-diazaspiro[5.5]undecal-9-ylmethyl)-2-fluorophenyl)ethanol (Example 67, step f) (0.13 g), 2-isobutyl a solution of the thiothiazole-4-carboxylic acid (Example 72, step d) (0.08 g) and triethylamine (0.23 mL) in DMF (7 mL) in 0 ° C 2h. The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The resulting gum was purified by silica gel chromatography eluting with 47.5:47.5:5 isohexane:ethyl acetate:triethylamine to 95:5 ethyl acetate:triethylamine. To the residue containing the product, toluene (200 mL) was evaporated. The yield was 0.24 g.

m/z 476(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.90 (s, 1H), 7.22 - 7.14 (m, 2H), 7.03 (t, J = 7.4 Hz, 1H), 4.35 (t, J = 4.7 Hz, 1H), 3.68-3.57 (m, 6H), 3.52-3.42 (m, 2H), 2.89 (d, J = 6.9 Hz, 2H), 2.76 (t, J = 7.5 Hz, 2H), 2.70 ( s, 2H), 2.45-2.28 (m, 4H), 2.12-2.03 (m, 1H), 1.74-1.65 (m, 2H), 1.57-1.48 (m, 2H), 0.96 (d, J = 6.7 Hz, 6H).

f) (R)-7-(2-(2-Fluoro-3-((4-(2-isobutylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

Adding trifluoroacetic acid (0.031 mL) to (9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undec carbon Alkyl-4-yl)(2-isobutylthiazol-4-yl)methanone (Example 72, step e) (0.19 g) in DCM (5 mL) The mixture was stirred for 5 min, then Dess-Martin periodinane (0.25 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (5mL), followed by addition of acetic acid (0.023 mL) and (R) -7- (2- amino-1-hydroxyethyl) -4-hydroxybenzo [d] thiazol -2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.13 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.038 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 35% acetonitrile). The title compound was obtained as a white solid. The yield was 0.10 g.

m/z 684(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.39-11.19 (m, 1H), 7.94 (s, 1H), 7.53-7.40 (m, 2H), 7.25 (t, J = 7.7Hz, 1H), 6.93 (d, J = 8.2 Hz, 1H), 6.77 (d, J = 8.2 Hz, 1H), 4.92 (dd, J = 8.5, 4.9 Hz, 1H), 4.33-4.26 (m, 2H), 3.72-3.59 (m, 6H), 3.28-3.01 (m, 10H), 2.88 (d, J = 6.9 Hz, 2H), 2.11-1.95 (m, 3H), 1.84-1.69 (m, 2H), 0.95 ( d, J = 6.7 Hz, 6H). Five unobserved exchangeable protons.

Example 73

(R)-7-(2-(2-fluoro-3-((4-(5-isopropylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (9-(2-Fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(5 -isopropylthiophen-3-yl)methanone

Adding O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.2g) to 2-(3-(1- Oxa-4,9-diazaspiro[5.5]undecal-9-ylmethyl)-2-fluorophenyl)ethanol (Example 67, step f) (0.13 g), 5-isopropyl A solution of thiophene-2-carboxylic acid (0.07 g) and triethylamine (0.23 mL) in DMF (7 mL) EtOAc. The resulting yellow solution was allowed to warm to RT and stirred for 2 h. The mixture was partitioned between EtOAc (EtOAc) (EtOAc)EtOAc. The resulting gum was purified by silica gel chromatography eluting with 47.5:47.5:5 isohexane:ethyl acetate:triethylamine to 95:5 ethyl acetate:triethylamine. To the residue containing the product was added toluene (200 mL). The yield was 0.23 g.

m/z 461(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.22 - 7.15 (m, 3H), 7.03 (t, J = 7.4 Hz, 1H), 6.83 (d, J = 3.6 Hz, 1H), 4.35 (t, J = 5.0 Hz, 1H), 3.69-3.58 (m, 6H), 3.49 (s, 2H), 3.21-3.13 (m, 1H), 2.76 (t, J = 8.4 Hz, 2H), 2.70 ( s, 2H), 2.42 - 2.36 (m, 4H), 1.75-1.68 (m, 2H), 1.56-1.46 (m, 2H), 1.29 (d, J = 6.7 Hz, 6H)

b) (R)-7-(2-(2-Fluoro-3-((4-(5-isopropylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

Adding trifluoroacetic acid (0.031 mL) to (9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undec carbon Alkyl-4-yl)(5-isopropylthiophen-2-yl)methanone (Example 73, step a) (0.18 g) in DCM (5 mL) The mixture was stirred for 5 min, then Dess-Martin periodinane (0.25 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (5mL), followed by addition of acetic acid (0.023 mL) and (R) -7- (2- amino-1-hydroxyethyl) -4-hydroxybenzo [d] thiazol -2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.13 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.025 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The title compound was obtained as a white solid. The yield was 0.12 g.

m/z 669(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.26 (s, 1H), 7.54 - 7.40 (m, 2H), 7.28 - 7.21 (m, 2H), 6.94 (d, J = 8.5 Hz, 1H), 6.84 (dd, J = 3.6, 0.8 Hz, 1H), 6.78 (d, J = 8.2 Hz, 1H), 4.93 (dd, J = 8.5, 4.9 Hz, 1H), 4.34 (s, 2H), 3.73-3.64 (m, 4H), 3.54 (s, 2H), 3.29-3.03 (m, 11H), 2.07-1.98 (m, 2H), 1.85-1.73 (m, 2H), 1.29 (d, J = 6.9) Hz, 6H). Five unobserved exchangeable protons.

Example 74

(R)-7-(2-(2-fluoro-3-((4-(4-isopropylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 1-(4-isopropylthiophen-2-yl)ethanone

1-(Thien-2-yl)ethanone (5.4 mL) was added to an ice-cold suspension of aluminum chloride (33 g) in anhydrous chloroform (100 mL). Then 2-bromopropane (5.2 mL) was added dropwise over 5 min. The reaction was allowed to warm to RT and stirred overnight. The dark suspension was carefully poured onto ice and the mixture was stirred for 10 min. The layers were separated and the aqueous extracted with DCM (100 mL). The combined organic layers were washed with EtOAc EtOAc (EtOAc m. The residue was purified by EtOAc (EtOAc) eluting The fractions containing the product are combined and evaporated to give the sub-title compound as a pale yellow oil. The yield was 5.4 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.58 (d, J = 1.3 Hz, 1H), 3.01-2.92 (m, 1H), 2.55 (s, 3H), 1.27 (d, J = 6.9 Hz, 6H). A thiophene proton that is obscured by the CDCl ₃ signal ~7.26.

b) 4-isopropylthiophene-2-carboxylic acid

1-(4-Isopropylthiophen-2-yl)ethanone (Example 74, step a) (1 g) in 1,4-dioxane (10 mL) was carefully added to sodium hydroxide (1.19) g) A solution of sodium hypochlorite (8%, 50 mL) in 60 °C. The resulting mixture was heated to 75 ° C and stirred for 1 h. The reaction was cooled and the aqueous was washed with EtOAc EtOAc The aqueous phase was treated with aqueous sodium bisulfite (10%, 20 mL) and carefully acidified with concentrated hydrochloric acid. The resulting mixture was extracted with DCM (3×50 mL). The combined organic solution was dried over sodium sulfate, filtered and evaporated to give a yellow oil. Purification by preparative HPLC (Sunfire ^TM, gradient: 0.2% T FA in an aqueous solution of 5 to 95% acetonitrile). The fractions containing the product are combined, evaporated and dried under high vacuum to give a sub-title compound as a white solid. The yield is 0.5g.

¹ H NMR (300 MHz, D ₆ -DMSO) δ 12.95 (s, 1H), 7.65 (d, J = 1.5 Hz, 1H), 7.51-7.49 (m, 1H), 2.94 (seven peak, J = 6.9 Hz , 1H), 1.20 (d, J = 6.9 Hz, 6H).

c) (9-(2-Fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.51 undecane-4-yl) (4- Isopropylthiophen-2-yl)methanone

Adding O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.2g) to 2-(3-(1- Oxa-4,9-diazaspiro[5.5]undecal-9-ylmethyl)-2-fluorophenyl)ethanol (Example 67, step f) (0.13 g), 4-isopropyl A solution of thiophene-2-carboxylic acid (Example 74, step b) (0.07 g) and triethylamine (0.23 mL) in DMF (7 mL) EtOAc. The resulting yellow solution was allowed to warm to RT and stirred for 2 h. The mixture was partitioned between EtOAc (EtOAc)EtOAc. The resulting gum was purified by silica gel chromatography eluting with 47.5:47.5:5 isohexane:ethyl acetate:triethylamine to 95:5 ethyl acetate:triethylamine. To the residue containing the product, toluene (200 mL) was evaporated. The yield was 0.23 g.

m/z 461(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ7.30-7.14 (m, 4H), 7.05-7.00 (m, 1H), 4.35 (s, 1H), 3.68-3.58 (m, 6H), 3.51-3.47 (m, 2H), 2.98-2.92 (m, 1H), 2.75 (t, J = 7.3 Hz, 2H), 2.70 (s, 2H), 2.41-2.34 (m, 4H), 1.75-1.68 ( m, 2H), 1.56-1.47 (m, 2H), 1.21. (d, J = 6.9 Hz, 6H).

d) (R)-7-(2-(2-Fluoro-3-((4-(4-isopropylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-mercaptobenzo[d]thiazole-2(3H)-one di-trifluoroacetate

Adding trifluoroacetic acid (0.031 mL) to (9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undec carbon Alkyl-4-yl)(4-isopropylthiophen-2-yl)methanone (Example 74, step c) (0.18 g) in DCM (5 mL) The mixture was stirred for 5 min, then Dess-Martin periodinane (0.25 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (5mL), followed by addition of acetic acid (0.023 mL) and (R) -7- (2- amino-1-hydroxyethyl) -4-hydroxybenzo [d] thiazol -2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.13 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.025 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5 = 5: 0.5 to 89: 10: 1 DCM: methanol: '880' aqueous ammonia solution. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The title compound was obtained as a white solid. The yield was 0.13 g.

m/z 669(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.26 (s, 1H), 7.53-7.47 (m, 1H), 7.47-7.41 (m, 1H), 7.31 (s, 1H), 7.29 ( s, 1H), 7.25 (t, J = 7.6 Hz, 1H), 6.94 (d, J = 8.5 Hz, 1H), 6.78 (d, J = 8.2 Hz, 1H), 4.93 (dd, J = 8.3, 4.7 Hz, 1H), 4.35-4.31 (m, 2H), 3.73-3.62 (m, 4H), 3.54 (s, 2H), 3.26-3.03 (m, 10H), 2.94 (seven peak, J = 7 Hz, 1H) , 2.08-1.98 (m, 2H), 1.85-1.71 (m, 2H), 1.21. (d, J = 6.9 Hz, 6H). Five unobserved exchangeable protons.

Example 75

(R)-7-(2-(2-chloro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 3-(carboxymethyl)-4-chlorobenzoic acid

Potassium hydroxide (1.549 g) in water (15 mL) was added to a suspension of 4-chloro-3-(cyanomethyl)benzoic acid (2.07 g) in ethanol (15 mL). It was heated under reflux for 4 hours and then allowed to cool. The mixture was concentrated in vacuo to remove ethanol, then diluted with water and washed twice with ethyl acetate. The organic phase was discarded and the aqueous phase was acidified to pH 1 with concentrated hydrochloric acid and extracted twice with ethyl acetate. The combined extracts were washed with EtOAc EtOAc m. The yield was 2.06 g.

m/z 214(M ⁺ )(EI)

b) 2-(2-chloro-5-(hydroxymethyl)phenyl)ethanol

Borane-methylsulfide complex solution (2M in THF, 12.0 mL) was added 3-(carboxymethyl)-4-chlorobenzoic acid in portions over 3 min (Example 75, step a) (2.06 g) It was suspended in anhydrous THF (30 mL) at room temperature. The resulting effervescent thick suspension was stirred at room temperature for 1 hour and then heated to reflux for 1 hour. The cooled mixture was quenched by the addition of methanol in portions over 2 min (10 mL). The solution was stirred at room temperature for 30 minutes and then concentrated over EtOAc (EtOAc) The yield was 0.983 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.35 (d, J = 8.2Hz, 1H), 7.28 (d, J = 2.1Hz, 1H), 7.18 (dd, J = 8.2,2.1Hz, 1H), 4.65 (s, 2H), 3.89 (t, J = 6.7 Hz, 2H), 3.02 (t, J = 6.5 Hz, 2H). Two unobserved exchangeable protons.

c) 4-chloro-3-(2-hydroxyethyl)benzaldehyde

Manganese (IV) (1.00 g) was added to 2-(2-chloro-5-(hydroxymethyl)phenyl)ethanol (Example 75, step b) (0.205 g) in DCM (10 mL) The solution was formed and the resulting suspension was stirred overnight at room temperature. The mixture was then filtered through Celite and the filter pad was washed thoroughly with DCM. The filtrate and washings were concentrated in vacuo to give the subtitle compound as a colourless oil. The yield was 0.159 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) 69.97 (s, 1H), 7.81 (d, J = 2.0,1H), 7.70 (dd, J = 2.0,8.2,1H), 7.53 (t, J = 6.7,1H) , 3.94 (dd, J = 6.4, 11.6, 2H), 3.10 (t, J = 6.6, 2H), 1.46 (t, J = 5.2, 1H).

d) (9-(4-Chloro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl) (2 -isopropylthiazol-4-yl)methanone

(2-Isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (0.178 g) of a solution in NMP (2 mL). Then a solution of 4-chloro-3-(2-hydroxyethyl)benzaldehyde (Example 75, step c) (0.154 g) in NMP (3 mL) was added and the solution was stirred for one hour and then triethyl hydrazide Treated with sodium oxyborohydride (0.181 g) and stirred at rt overnight. More sodium triethoxysulfonium borohydride (0.404 g) was added, and the mixture was stirred for additional 6 hr then poured into saturated sodium bicarbonate and extracted twice with ethyl acetate. The combined extracts were washed three times with water, once with brine, then dried over anhydrous magnesium sulfate and purified by flash chromatography on ruthenium dioxide, eluting with 1:2:97 triethylamine:methanol:dichloromethane, a colorless gum of the sub-title compound. The yield was 0.157 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.90 (s, 1H), 7.29 (d, J = 8.2 Hz, 1H), 7.24 (d, J = 1.4 Hz, 1H), 7.11 (dd , J=8.2, 1.8 Hz, 1H), 4.37 (t, J = 5.3 Hz, 1H), 3.73-3.56 (m, 8H), 3.41 (s, 2H), 3.35-3.28 (m, 1H), 2.85 ( t, J = 6.9 Hz, 2H), 2.41-2.10 (m, 4H), 1.75-1.62 (m, 2H), 1.60-1.46 (m, 2H), 1.36 (d, J = 6.7 Hz, 6H).

e) 2-(2-chloro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.51 undecane-9- Methyl)phenyl)acetaldehyde

(9-(4-Chloro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-iso A solution of the propyl thiazol-4-yl)methanone (Example 75, step d) (0.152 g) in EtOAc (EtOAc) Dess-Martin periodinane (0.205 g) was added, after which the mixture was taken out from the cooling bath and stirred at room temperature for 30 minutes. The solution was diluted with a saturated aqueous solution of sodium thiosulfate (5 mL), saturated sodium bicarbonate (5 mL) and ethyl acetate (5 mL), and the mixture was stirred vigorously for 10 min. The mixture was then extracted with EtOAc (EtOAc)EtOAc. The yield was 0.197 g.

m/z 476(M+H) ⁺ (APCI)

f) (R)-7-(2-(2-chloro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

( R )-7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) A solution of 0.145 g) in MeOH (2 mL) wasEtOAc (EtOAc) Subsequent addition of 2-(2-chloro-5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 a solution of -methyl)phenyl)acetaldehyde (Example 75, step e) (0.197 g) in MeOH (3 mL), and the resulting mixture was stirred at room temperature for 5 min then cooled in ice-water. Treated with sodium cyanoborohydride (0.039 g). The cooling bath was removed and the mixture was stirred at room temperature for 140 min then treated with more sodium cyanoborohydride (0.040 g). The mixture was stirred overnight. Next morning, the mixture was quenched to a drop of water, the solution was filtered and purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in acetonitrile 15 to 35%) by. The product containing fractions were concentrated in vacuo and co-evaporated three times from acetonitrile to give a colourless residue. The residue was triturated with EtOAc (EtOAc)EtOAc. The yield was 0.077 g.

m/z 686/688(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.94 (s, 1H), 7.53 (d, J = 8.2 Hz, 1H), 7.48-7.39 (m, 2H), 6.94 (d, J = 8.2 Hz, 1H), 6.77 (d, J = 8.5 Hz, 1H), 4.91 (dd, J = 8.5, 4.9 Hz, 1H), 4.22 (s, 2H), 3.75-3.60 (m, 6H), 3.52- 2.96 (m, 11H), 2.09-1.90 (m, 2H), 1.83-1.62 (m, 2H), 1.35 (d, J = 6.9 Hz, 6H). Six unobserved exchangeable protons.

Example 76

(R)-8-hydroxy-5-(1-hydroxy-2-(3-((4-(4-isopropylthiazol-2-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)methyl)phenethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

a)(R)-5-(1-( third -butylisomethyl decyloxy)-2-(3-((4-(4-isopropylthiazol-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Undecane-9-yl)methyl)phenethylamino)ethyl)-8-hydroxyquinoline-2(1H)-one

(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(4-isopropylthiazole- A solution of 2-yl)methanone (Example 66, step c) (0.195 g) in EtOAc (EtOAc) (EtOAc) The resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.025 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3mL) and added to (R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8-hydroxy-quinolin-2 (1H)-ketone (WO 2004106333) (191 mg) in MeOH (15 mL). The mixture was cooled to 0 ° C and sodium triacetoxyborohydride (0.140 g) was added in one portion. The reaction mixture was stirred at 20 ° C for 3 hours. The solvent was removed under reduced pressure and the residue was partitioned between ethyl acetate and saturated sodium hydrogen carbonate. The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The crude product was purified by flash chromatography on EtOAc (EtOAc) eluting EtOAc EtOAc The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.133 g.

m/z 760(M+H) ⁺ (APCI)

b) (R)-8-hydroxy-5-(1-hydroxy-2-(3-((4-(4-isopropylthiazole-2-carbonyl)-1-oxa-4,9-diaza) Heterospiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

In methanol (1 mL) of the triethylamine trihydrofluoride (0.034 mL) was added to (R) -5- (1- (third - butyldimethylsilyl silicon alkyloxy) -2- (3- ((4-(4-Isopropylthiazole-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl)phenethylamino Ethyl)-8-hydroxyquinolin-2(1H)-one (Example 76, step a) (0.133 g) in THF (4 mL). The solvent was removed under reduced pressure and the crude product was purified by preparative HPLC (Sunfire ^(TM) , gradient: 10 to 40% acetonitrile in 0.2% aqueous TFA). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.090 g.

m/z 646(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D _6- DMSO, 90 ° C) δ 8.17 (d, J = 10.0 Hz, 1H), 7.51 (s, 1H), 7.44 - 7.33 (m, 4H), 7.14 (d, J = 8.2 Hz, 1H), 7.00 (d, J = 8.2 Hz, 1H), 6.54 (d, J = 10.0 Hz, 1H), 5.38-5.33 (m, 1H), 4.28 (s, 2H), 3.78-3.74 ( m, 2H), 4.13 - 3.47 (m, 4H), 3.28 (t, J = 8.2 Hz, 2H), 3.21-3.00 (m, 9H), 2.09-1.99 (m, 2H), 1.84-1.72 (m, 2H), 1.26 (d, J = 6.8 Hz, 6H). Six unobserved exchangeable protons.

Example 77

(R)-5-(2-(2,3-difluoro-4-(2-(4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)ethoxy)phenethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetate

a) 2-(2,3-difluoro-4-hydroxyphenyl)acetic acid

Boron tribromide solution (1M in DCM, 13.4 mL) was added dropwise to 2-(2,3-difluoro-4-methoxyphenyl)acetic acid (1.18 g) in DCM (5 mL) In a suspension of °C. The reaction was allowed to warm to RT and stirred overnight. The reaction was cooled to -78.degree. C. and a boron tribromide solution (1M in DCM, 13.4 mL). The reaction was allowed to warm to RT and stirred for 1 h. Pour the reaction on ice. The resulting aqueous solution was extracted with DCM (5×50 mL). The aqueous phase was then extracted with ethyl acetate (3×100 mL). The combined organic solution was washed with EtOAc EtOAc m. The yield was 1.09 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ12.43 (s, 1H), 10.24 (s, 1H), 6.91 (td, J = 8.3,2.1Hz, 1H), 6.72 (td, J = 8.3,1.8 Hz, 1H), 3.54 (s, 2H).

b) 2,3-difluoro-4-(2-hydroxyethyl)phenol

A borane dimethyl sulfide solution complex (2M in THF, 14.4 mL) was added dropwise to 2-(2,3-difluoro-4-hydroxyphenyl)acetic acid (Example 77, step a) 1.08 g) The resulting mixture was warmed to RT and stirred overnight in tetrahydrofuran (25 mL) EtOAc. The reaction was stopped with methanol and the solvent was evaporated when bubbling ceased. The residue was purified by EtOAc (EtOAc) elute The fractions containing the product are combined and evaporated to give the subtitle compound as a white solid. The yield was 0.95 g.

¹ H NMR (300 MHz, D ₆ -DMSO) δ 10.08 (s, 1H), 6.87 (td, J = 8.3, 2.2 Hz, 1H), 6.69 (td, J = 8.4, 1.9 Hz, 1H), 4.68 ( t, J = 5.3 Hz, 1H), 3.58-3.49 (m, 2H), 2.66 (t, J = 7.0 Hz, 2H). c) 2-(4-(2,2-diethoxyethoxy)-2,3-difluorophenyl)ethanol

Barium carbonate (0.99 g) was added to 2,3-difluoro-4-(2-hydroxyethyl)phenol (Example 77, step b) (0.44 g) and 2-bromo-1,1-diethoxy A solution of ethane (0.4 mL) in DMF (10 mL). The resulting suspension was heated at 90 ° C for 18 h. The reaction was cooled and poured into water (100 mL). The aqueous mixture was extracted with diethyl ether (3 x 100 mL). The combined organic solution was washed with water (200 mL) brine brine The crude material was purified by EtOAc (EtOAc) elute The fractions containing the product are combined and evaporated to give the sub-title compound as a yellow oil. The yield was 0.49 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 7.06 - 6.93 (m, 2H), 4.81 (t, J = 5.1 Hz, 1H), 4.70 (t, J = 5.3 Hz, 1H), 4.02 (d, J = 5.4 Hz, 2H), 3.73 - 3.62 (m, 2H), 3.61-3.50 (m, 4H), 2.71 (t, J = 6.9 Hz, 2H), 1.13 (t, J = 7.0 Hz, 6H).

d) 2-(2,3-difluoro-4-(2-hydroxyethyl)phenoxy)acetaldehyde

Concentrated hydrochloric acid (5 mL) was added to 2-(4-(2,2-diethoxyethoxy)-2,3-difluorophenyl)ethanol (Example 77, step c) (0.49 g) The solution in 1,4-dioxane (10 mL) and the resulting mixture was stirred for 1 h. The mixture was carefully diluted with water (50 mL) and ethyl acetate (3×50 mL). The combined organics were washed with water (50 mL) EtOAc. The yield was 0.31 g.

e) (9-(2-(2,3-difluoro-4-(2-hydroxyethyl)phenoxy)ethyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkan-4-yl)(2-isopropylthiazol-4-yl)methanone

(2-Isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, Step b) (0.5 g) was added to 2-(2,3-difluoro-4-(2-hydroxyethyl)phenoxy)acetaldehyde (Example 77, step d) (0.31 g) in N-A In a solution of keto-pyrrolidone (10 mL) and acetic acid (0.07 mL). The resulting mixture was stirred for 30 min and then cooled in an ice bath. Sodium triethoxy borohydride (0.38 g) was then added and the reaction allowed to warm to RT and stirred overnight. The reaction mixture was partitioned between ethyl acetate (100 mL) and water (100 mL). The aqueous phase was basified and saturated with saturated sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (2×100 mL). The combined organic solution was washed with water (100 mL) brine brine The residue was purified by silica gel chromatography eluting EtOAc EtOAc:EtOAc:EtOAc The product-containing fractions are combined and evaporated to give a clear gel of the sub-title compound. The yield was 0.41 g.

m/z 510(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ7.93 (s, 1H), 7.06-6.85 (m, 2H), 4.58-4.36 (m, 1H), 4.13 (t, J = 5.7Hz, 2H), 3.73-3.51 (m, 8H), 3.36-3.23 (m, 1H), 2.76-2.63 (m, 4H), 1.76-1.45 (m, 4H), 1.35 (d, J = 6.9 Hz, 6H) +4 protons blurred by DMSO peaks.

f) (R)-5-(2-(2,3-difluoro-4-(2-(4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)ethoxy)phenethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetic acid salt

Adding trifluoroacetic acid (0.03 mL) to (9-(2-(2,3-difluoro-4-(2-hydroxyethyl)phenoxy)ethyl)-1-oxa-4,9- Diazaspiro[5.5]undecethane-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 77, step e) (0.18 g) in DCM (5 mL) In a solution of °C. The reaction was stirred for 5 min, then Dess-Martin periodinane (0.225 g) was added. The mixture was allowed to warm to RT and stirred for 1 h. Saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL). The layers were separated and the aqueous was extracted with ethyl acetate (20 mL). The combined organics were washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was redissolved in methanol (5mL), acetic acid (0.02 mL) and (R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8 -Hydroxyquinolin-2(1H)-one (WO 2004106333) (0.12 g), the mixture was stirred for 5 min and cooled in an ice bath. Sodium cyanoborohydride (0.033 g) was then added, which was allowed to warm to RT and stirred overnight. The reaction was concentrated and purified by silica gel chromatography eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM: methanol: &lt; The fractions containing the product are combined and evaporated. The residue was dissolved in THF (5 mL) EtOAc (EtOAc) The solvent was evaporated and the residue was azeotroped twice with toluene. Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 10 to 30% acetonitrile). The title compound was obtained as a white solid. The yield was 0.11 g.

m/z 712(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.18 (d, J = 9.7 Hz, 1H), 7.96 (s, 1H), 7.18-6.96 (m, 4H), 6.54 (d, J = 10.0 Hz, 1H), 5.36 (dd, J=8.7, 4.1 Hz, 1H), 4.50-4.42 (m, 2H), 3.76-3.65 (m, 6H), 3.62-3.54 (m, 2H), 3.44-2.98 (m, 11H), 2.12-2.01 (m, 2H), 1.90-1.75 (m, 2H), 1.35 (d, J = 6.7 Hz, 6H). Six unobserved exchangeable protons.

Example 78

(R)-7-(2-(3-((4-(2-butylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)methyl)-2-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) third -butyl(2-fluorophenylethoxy)dimethyl decane

Third - butyldimethylsilyl silicon alkyl chloride (6.45 g of) was added to 2- (2-fluorophenyl) ethanol (5.00 g) and 1H- imidazole (7.29g) in anhydrous DMF (30mL) in an ice bath The medium is cooled in a stirred solution. After 45 min the reaction mixture was diluted with EtOAc EtOAc. The resulting gum was dissolved in isohexane and applied to a ruthenium tube column, eluted with isohexane, followed by 1:3 ethyl acetate: isohexane to collect the product oil. The yield was 9.0 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 7.28-7.18 (m, 2H), 7.13-7.00 (m, 2H), 3.84 (t, J = 6.8Hz, 2H), 2.89 (t, J = 7.2Hz, 2H ), 0.89 (s, 9H), 0.008 (s, 6H).

b) 3-(2-(Third-butyldimethylammoniooxy)ethyl)-2-fluorobenzaldehyde

Tri-butyl(2-fluorophenylethoxy)dimethyl decane (Example 78, step a) (5.00 g) was added over 5 min to a second butyl lithium (1.4 M solution in cyclohexane, 14.0 mL) and N1 - (2- (dimethylamino) ethyl) - N1, N2, N2 - trimethyl-1,2-diamine (4.10 mL) in THF (25mL) was cooled to -78 ° C in a stirred solution. After 2 h, N,N -dimethylformamide (10.06 g) was added and the reaction mixture was stirred at -78 °C for 1 h and then cooled. After another 0, 5 h, the reaction was stopped in water. Ethyl acetate (300 mL) was added, and the mixture was washed with water (3.times.150 mL), 2M HCl (2×50 mL), water (2×50 mL), brine, dried over sodium sulfate An oil that produces a sub-title compound. The yield was 5.3 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ10.40 (s, 1H), 7.76 (dt, J = 2.0 and 6.8Hz, 1H), 7.54 (dt , J = 1.6 and 7.2Hz, 1H), 7.21 (t , J = 7.2 Hz, 1H), 3.88 (t, J = 5.2 Hz, 2H), 2.95 (dt, J = 1.2 and 6.4 Hz, 2H), 0.88 (s, 9H), 0.008 (s, 6H).

c) 1-(9-(3-(2-( third -butyldimethylmethylalkyloxy)ethyl)-2-fluorobenzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)-2, 2,2-trifluoroethyl ketone

3-(2-( Third -butyldimethylsulfonyloxy)ethyl)-2-fluorobenzaldehyde (Example 78, step b) (2.159 g) was added to 2,2,2-tri Fluor-1-(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)ethanone trifluoroacetate (Example 12, step d) (2.80 g) and acetic acid (0.438 mL) in a stirred solution of NMP (25 mL). After 5 minutes, sodium triethoxysulfonate hydride (3.24 g) was added. After 16 h, water was added and the mixture was partitioned between water (250 mL) and ethyl acetate (250 mL). The ethyl acetate solution was washed with water (3 x 250 mL) and brine then evaporated. Purification by silica gel chromatography, 20:1:1, isohexane:ethyl acetate: triethylamine eluted to give a sub-title compound. The yield is 2.5g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.27-7.20 (m, 1H), 7.20-7.13 (m, 1H), 7.08-7.01 (m, 1H), 3.84 (t, J = 7.2Hz, 3H), 3.80-3.77 (m, 2H), 3.71-3.67 (m, 1H), 3.64-3.58 (m, 3H), 3.56 (s, 1H), 3.41 (s, 1H), 2.90 (q, J = 6.4 Hz, 2H), 2.69-2.62 (m, 1H), 2.52-2.44 (m, 1H), 2.43-2.35 (m, 1H), 1.92-1.80 (m, 2H), 1.72-1.60 (m, 2H), 0.89 ( s, 9H), 0.00 (s, 6H).

d) 9-(3-(2-(Third-butyl dimethyl decyloxy)ethyl)-2-fluorobenzyl)-1-oxa-4,9-diazaspiro[5.51 Undecane

The '880' aqueous ammonia solution (3.0 mL) was added to 1- (9- (3- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethyl) -2-fluoro-benzyl) -1 Oxa-4,9-diazaspiro[5.5]undecyl-4-yl)-2,2,2-trifluoroethanone (Example 78, step c) (2.5 g) in MeOH (10 mL) In the stirred solution. After 1 h, the reaction mixture was evaporated to dryness. Acetonitrile is added and the solution is evaporated to dryness in vacuo. This procedure is repeated three times to give an oil of sub-title compound. The yield was 2.07 g. Use directly.

m/z 423(M+H) ⁺ (APCI)

e) pentamidine

Phosphorus pentasulfide (5.56 g) was added to a stirred suspension of pentaneguanamine (10.0 g) in methyl tri -butyl ether (300 mL). After 16 h, the mixture was filtered with EtOAc EtOAc (EtOAc) The yield was 11.4 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 9.35-9.23 (br s, 1H), 9.15-9.05 (br s, 1H), 2.52-2.42 (m, 2H), 1.68-1.55 (m, 2H) , 1.35 - 1.22 (m, 2H), 0.87 (t, J = 7.6 Hz, 3H).

f) ethyl 2-butylthiazole-4-carboxylate

Ethyl 3-bromo-2-oxopropionate (6.76 mL) was added with great care to a stirred solution of pentanethioindole (Example 78, step e) (6.30 g) in ethanol (100 mL). The solution was then heated under reflux for 16 h. After cooling, the reaction mixture was diluted with EtOAc EtOAc. Purification by chromatography on silica gel eluting with EtOAc (EtOAc:EtOAc) The yield was 6.5 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ8.05 (s, 1H), 4.42 (q, J = 7.1Hz, 2H), 3.02-3.05 (m, 2H), 1.85-1.75 (m, 2H), 1.50- 1.38 (m, 5H), 0.95 (t, J = 7.1 Hz, 3H).

g ) 2-butylthiazole-4-carboxylic acid

To a stirred mixture of THF (80 mL) and water (20 mL), EtOAc (EtOAc) After 16 h, concentrated hydrochloric acid (10 mL) was added and the solution was concentrated to ~40 mL. The reaction mixture was partitioned between ethyl acetate and brine. The aqueous layer was extracted three times with ethyl acetate. The combined organic layers were dried with MgSO4, filtered and evaporated The yield was 4.6 g.

¹ H NMR (300MHz, CDCl ₃ ) δ 8.18 (s, 1H), 3.11-3.03 (m, 2H), 1.88-1.78 (m, 2H), 1.52-1.38 (m, 2H), 0.97 (t, J) = 7.2 Hz, 3H). An unobserved exchangeable proton.

h)(9-(3-(2-( third -butyldimethylmethylalkyloxy)ethyl)-2-fluorobenzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2- Butylthiazole-4-yl)methanone

2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethylisourea ruthenium hexafluorophosphate (V) ) (0.378 g) was added to 2-butyl-4-carboxylic acid (Example 78, step g) (0.193g), 9- ( 3- (2- ( tertiary - butyl dimethyl silicone alkyloxy Ethyl)-2-fluorobenzyl)-1-oxa-4,9-diazaspiro[5.5]undecane (Example 78, step d) (0.4 g) and triethylamine (0.383) g) in a stirred solution in DMF (4 mL). After 1 h, the reaction mixture was partitioned between water and ethyl acetate. The ethyl acetate layer was washed twice with water and brine, dried over magnesium sulfate, filtered and evaporated. Purification by silica gel chromatography, eluting with 7:1:0.5 isohexane:ethyl acetate:triethylamine afforded the subtitle compound. The yield was 0.35 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ7.94 (s, 1H), 7.30-7.16 (m, 2H), 7.13-7.05 (m, 1H), 3.88-3.80 (m, 2H), 3.74-3.62 (m, 6H), 3.55-3.49 (m, 2H), 3.09-3.00 (m, 2H under water peak), 2.87-2.79 (m, 2H), 2.49-2.32 (m, 4H), 1.84 -1.68 (m, 4H), 1.62-1.51 (m, 2H), 1.50-1.39 (m, 2H), 0.97 (t, J = 7.9 Hz, 3H), 0.87 (s, 9H), 0.00 (s, 6H) ).

i) (2-butylthiazol-4-yl)(9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] Undecyl-4-yl)methanone

The TBAF (1M solution in THF, 2.0mL) was added to (9- (3- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethyl) -2-fluoro-benzyl) -1 Oxa-4,9-diazaspiro[5.5]undecethane-4-yl)(2-butylthiazol-4-yl)methanone (Example 78, step h) (0.350 g) in THF In a solution (4 mL). After 0.5 h, the solution was concentrated. Purification by silica gel chromatography eluting with 10:1 ethyl acetate: triethylamine afforded the subtitle compound. The yield was 0.24 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.88 (s, 1H), 7.22 - 7.13 (m, 2H), 7.03 (t, J = 7.6 Hz, 1H), 4.35 (t, J = 5.2 Hz, 1H), 3.68-3.56 (m, 12H), 3.48 (s, 2H), 3.00 (t, J = 7.5 Hz, 2H), 2.75 (t, J = 7.5 Hz, 2H), 2.44 - 2.28 ( m, 4H), 1.78-1.65 (m, 2H), 1.45-1.33 (m, 2H), 0.91 (t, J = 7.5 Hz, 3H).

j) 2-(3-((4-(2-butylthiazole-4-carbonyl)-4-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl )-2-fluorophenyl)acetaldehyde

Add Dess-Martin periodinane (0.278 g) to (2-butylthiazol-4-yl)(9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa -4,9-diazaspiro[5.5]undecane-4-yl)methanone (Example 78, step i) (0.240 g) and trifluoroacetic acid (0.058 mL) in DCM (5 mL) Stir the solution. After 1 h, ethyl acetate (30 mL) was added followed by a mixture of saturated sodium thiosulfate (5 mL) and saturated sodium bicarbonate (5 mL). The reaction mixture was thoroughly shaken and separated. The ethyl acetate solution was washed with saturated sodium bicarbonate solution, water and brine. Acetic acid (0.08 mL) was added, the solution was dried over sodium sulfate, filtered and evaporated in vacuo. The yield was 0.24 g. Use directly.

m/z 474(M+H) ⁺ (APCI)

k)( R )-7-(2-(3-((4-(2-butylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane) -9-yl)methyl)-2-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

Acetic acid (0.044 mL) was added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.200 g) and 2-(3-((4-(2-butylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] To a stirred solution of hexanes (10 mL) in MeOH (10 mL). After 1 min, sodium cyanoborohydride (0.064 g) was added. After 1.5h, the reaction mixture was filtered and purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in acetonitrile 10 to 40%) by. The fractions containing the pure product were combined and evaporated in vacuo. Acetonitrile (200 mL) was added and the solution was evaporated in vacuo to a gum. Repeat this process twice. Diethyl ether was added and the title compound was collected as a solid. The yield was 0.15 g.

m/z 684(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.28 (br s, 1H), 7.92 (s, 1H), 7.52-7.37 (m, 2H), 7.25 (t, J = 8.4Hz, 1H ), 6.93 (d, J = 8.3 Hz, 1H), 6.77 (d, J = 8.3 Hz, 1H), 4.94 - 4.87 (m, 1H), 4.30 (s, 2H), 3.70 (br s, 6H), 3.63 (s, 2H), 3.28-2.92 (m, 10H), 2.06-1.94 (m, 2H), 1.82-1.66 (m, 4H), 1.43-1.31 (m, 2H), 0.90 (t, J = 7.6) Hz, 3H). Five unobserved exchangeable protons.

Example 79

(R)-7-(2-(3-((4-(benzo[b]thiophen-2-yl)-1-oxa-4,9-diazaspiro[5.5]undecane- 9-yl)methyl)-2-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzene well [d]thiazole-2(3H)-one di-trifluoroacetate

a) benzo[b]thiophen-2-yl (9-(3-(2-) third -butyldimethylmethylalkyloxy)ethyl)-2-fluorobenzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone

2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethylisourea ruthenium hexafluorophosphate (V) ) (0.378 g) was added to the benzo [b] thiophene-2-carboxylic acid (0.186g), 9- (3- ( 2- ( tertiary - butyl dimethyl silicone alkyloxy) ethyl) -2- Fluorobenzyl)-1-oxa-4,9-diazaspiro[5.5]undecane (Example 78, step d) (0.40 g) and triethylamine (0.383 g) in DMF (4 mL) In the stirred solution. After 1 h, the reaction mixture was partitioned between water and ethyl acetate. The ethyl acetate layer was washed twice with water and brine brine and dried over magnesium sulfate. Purification by silica gel chromatography, eluting with 7:1:0.5 isohexane:ethyl acetate:triethylamine afforded the subtitle compound. The yield was 0.38 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 8.14 - 8.08 (m, 1 H), 8.05-7.99 (m, 1H), 7.82 (s, 1H), 7.57-7.51 (m, 2H), 7.33-7.23 (m, 2H), 7.14 (t, J = 8.1 Hz, 1H), 3.86 (t, J = 7.3 Hz, 2H), 3.81-3.72 (m, 4H), 3.63 (s, 2H), 3.56 (s, 2H), 3.38 (s, 2H), 2.86 (t, J = 7.4 Hz, 2H), 2.46-2.34 (m, 2H), 1.89-1.81 (m, 2H), 1.66-1.52 (m, 2H), 0.88 (s, 9H), 0.00 (s, 6H).

b) benzene well [b]thiophen-2-yl (9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkan-4-yl)methanone

The TBAF (1M solution in THF, 2mL) was added to the benzo [b] thiophen-2-yl (9- (3- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethyl) - 2-fluorobenzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone (Example 79, step a) (0.370 g) in THF (4 mL In the solution. After 0.5 h, the solution was concentrated. Purification by silica gel chromatography eluting with 10:1 ethyl acetate: triethylamine afforded the subtitle compound. The yield was 0.26 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ7.99-7.95 (m, 1H), 7.93-7.88 (m, 1H), 7.67 (s, 1H), 7.46-7.39 (m, 2H), 7.17 (q, J = 7.5 Hz, 2H), 7.01 (t, J = 8.0 Hz, 1H), 4.39 - 4.33 (m, 1H), 3.72-3.57 (m, 4H), 3.54 (s, 2H), 3.48 (s, 2H), 2.99 (s, 2H), 2.75 (t, J = 7.6 Hz, 2H), 2.46-2.32 (m, 4H), 1, 80-1.71 (m, 2H), 1.58-1.48 (m , 2H).

c) 2-(3-((4-(Benzo[b]thiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)) Base)-2-fluorophenyl)acetaldehyde

Add Dess-Martin periodine (0.282 g) to benzo[b]thiophen-2-yl(9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa- Stirring of 4,9-diazaspiro[5.5]undec-4-yl)methanone (Example 79, step b) (0.240 g) and trifluoroacetic acid (0.059 mL) in DCM (5 mL) Solution. After 1 h, ethyl acetate (30 mL) was added followed by a mixture of saturated sodium thiosulfate (5 mL) and saturated sodium bicarbonate (5 mL). The reaction mixture was thoroughly shaken and separated. The ethyl acetate solution was washed with saturated sodium bicarbonate solution, water and brine. Acetic acid (0.08 mL) was added, then the solution was dried over sodium sulfate, filtered and evaporated in vacuo (j. The yield was 0.24 g. Use directly.

m/z 467(M+H) ⁺ (APCI)

d) (R)-7-(2-(3-((4-(benzo[b]thiophen-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecene Alkan-9-yl)methyl)-2-fluorophenethylamino)-1-indolylethyl)-4-mercaptobenzo[d]thiazole-2(3H)-one di-trifluoroacetic acid salt

Acetic acid (0.044 mL) was added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.203 g) and 2-(3-((4-(benzo[b]thiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5] A stirred solution of undecane-9-yl)methyl)-2-fluorophenyl)acetaldehyde (Example 79, step c) (0.240 g) in MeOH (10 mL). After 1 min, sodium cyanoborohydride (0.081 g) was added. After 1.5h, the reaction mixture was filtered and purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in acetonitrile 10 to 40%) by. The fractions containing the pure product were combined and evaporated in vacuo. Acetonitrile (200 mL) was added and the solution was evaporated in vacuo to a gum. Repeat this process twice. Diethyl ether was added and the title compound was collected as a solid. The yield was 0.19 g.

m/z 677(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.28 (s, 1H), 8.00-7.95 (m, 1H), 7.92-7.88 (m, 1H), 7.69 (s, 1H), 7.50- 7.38 (m, 4H), 7.25 (t, J = 7.5 Hz, 1H), 6.94 (d, J = 8.4 Hz, 1H), 6.77 (d, J = 8.4 Hz, 1H), 4.90 (dd, J = 4.2) And 8.56 Hz, 1H), 4.35-4.19 (m, 2H), 3.76-3.68 (m, 4H), 3.59 (s, 2H), 3.27-2.99 (m, 10H), 2.09-1.99 (m, 2H), 1.82-1.70 (m, 2H). Five unobserved exchangeable protons.

Example 80

(R)-7-(2-(2-chloro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-Chloro-3-(cyanomethyl)benzoic acid

A solution of 3-(bromomethyl)-2-chlorobenzoic acid (6.39 g) in DMF (75 mL)EtOAc. Overnight. The mixture was diluted with water and extracted twice with ethyl acetate. The organic phase was discarded and the aqueous phase was carefully acidified (25 mL) with concentrated hydrochloric acid to remove any HCN that was evolved from the bleaching solution. After stirring for two hours, the aqueous phase was extracted twice more with ethyl acetate. The combined organic phase was washed with EtOAc (EtOAc)EtOAc. The yield was 4.12 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 7.73 (dd, J = 7.7, 1.5 Hz, 1H), 7.69 (dd, J = 7.8, 1.7 Hz, 1H), 7.48 (t, J = 7.7 Hz, 1H), 4.16 (s, 2H) + 1 unobserved exchangeable protons.

b) 3-(carboxymethyl)-2-chlorobenzoic acid

Potassium hydroxide (2.976g) in water (30mL) was added to 2-chloro-3-(cyanomethyl)benzoic acid (Example 80, step a) (4.12g) in ethanol (30mL) The suspension was formed and the resulting solution was heated under reflux for 2 hours and then allowed to cool overnight. The mixture was concentrated in vacuo to remove ethanol, then diluted with water and washed twice with ethyl acetate. The organic phase was discarded and the aqueous phase was acidified to pH 1 with concentrated hydrochloric acid and extracted three times with ethyl acetate. The combined extracts were dried with EtOAc EtOAc m. The yield was 4.11 g. Use directly.

c) 2-(2-chloro-3-(hydroxymethyl)phenyl)ethanol

Borane-methylthio complex solution (2M in THF, 20 mL) was added portionwise to 3-(carboxymethyl)-2-chlorobenzoic acid in 5 min (Example 80, step b) (4.11 g) In a suspension of anhydrous THF (100 mL) at room temperature. The resulting effervescent suspension was stirred at room temperature for 30 minutes, then heated to reflux for 60 minutes and allowed to cool to room temperature overnight. The mixture was quenched by the addition of methanol in 15 minutes to quench the reaction (15 mL). The mixture was further diluted with methanol to give a solution which was then concentrated in vacuo to give a syrup. The syrup was purified by flash chromatography on EtOAc (EtOAc) elute The yield was 1.44 g.

m/z 186(M ⁺ )(EI)

d) 2-chloro-3-(2-hydroxyethyl)benzaldehyde

Manganese (IV) (1.599 g) was added to 2-(2-chloro-3-(hydroxymethyl)phenyl)ethanol (Example 80, step c) (0.342 g) in DCM (20 mL) The solution was formed and the resulting suspension was stirred overnight at room temperature. The mixture was then concentrated on EtOAc (EtOAc m.). The yield was 0.223 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ10.54 (d, J = 0.8Hz, 1H), 7.83 (dd, J = 7.7,1.5Hz, 1H), 7.55 (dd, J = 7.7,1.8Hz, 1H) , 7.35 (t, J = 7.6 Hz, 1H), 3.94 (dd, J = 11.0, 6.4 Hz, 2H), 3.11 (t, J = 6.6 Hz, 2H), 1.46 (t, J = 4.9 Hz, 1H) .

e) (9-(2-Chloro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -isopropylthiazol-4-yl)methanone

(2-Isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (0.325 g) of a solution in NMP (3 mL). Subsequently, a solution of 2-chloro-3-(2-hydroxyethyl)benzaldehyde (Example 80, step d) (0.217 g) in NMP (4 mL) was added, and the resulting solution was stirred for 1 hour, and then with three Treatment with sodium acetoxyborohydride (1.025 g). The mixture was stirred at room temperature overnight then poured into saturated sodium hydrogen The combined extracts were washed three times with water, once with brine, then dried over anhydrous magnesium sulfate and purified by flash chromatography on ruthenium dioxide, eluting with 1:2:97 triethylamine:methanol:dichloromethane, a colorless gum of the sub-title compound. The yield was 0.411 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.91 (d, J = 1.3 Hz, 1H), 7.30 (d, J = 7.2 Hz, 1H), 7.24 - 7.16 (m, 2H), 4.36 (t, J = 5.1 Hz, 1H), 3.73 - 3.59 (m, 8H), 3.55 (s, 2H), 3.32 (seven peak, J = 6.8 Hz, 1H), 2.89 (t, J = 7.0 Hz, 2H) ), 2.46-2.40 (m, 1H), 2.40-2.28 (m, 1H), 2.17 (t, J = 8.1 Hz, 1H), 1.91 (five peaks, J = 7.5 Hz, 1H), 1.77-1.65 ( m, 2H), 1.62-1.50 (m, 2H), 1.36 (d, J = 6.5 Hz, 6H).

f) 2-(2-Chloro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)methyl)phenyl)acetaldehyde

(9-(2-Chloro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-iso A solution of propyl thiazol-4-yl)methanone (Example 80, step e) (0.386 g) in EtOAc (EtOAc)EtOAc. Dess-Martin periodinane (0.519 g) was added, after which the mixture was taken out from the cooling bath and stirred at room temperature for 35 minutes. The solution was diluted with a saturated aqueous solution of sodium thiosulfate (5 mL), saturated sodium bicarbonate (5 mL) and ethyl acetate (5 mL), and the mixture was stirred vigorously for 10 min. The mixture was extracted with EtOAc (EtOAc) EtOAc. . The yield was 0.494 g.

m/z 476(M+H) ⁺ (APCI)

g) (R)-7-(2-(2-Chloro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

( R )-7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) A solution of 0.181 g) in MeOH (2 mL) wasEtOAc (EtOAc) Subsequent addition of 2-(2-chloro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 a solution of -methyl)phenyl)acetaldehyde (Example 80, step f) (0.247 g) in MeOH (3 mL), and the mixture was stirred at room temperature for 10 min then cooled in ice-water. Treated with sodium cyanoborohydride (0.102 g). The cooling bath was removed and the mixture was stirred at room temperature overnight. The next day, more sodium cyanoborohydride (0.099 g) was added and the mixture was stirred for another 4 hours. It was quenched by adding drop of water, filtered and purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile) to borrow. The product containing fractions were concentrated in vacuo and co-evaporated three times from acetonitrile to give a colourless residue. The residue was triturated with EtOAc (EtOAc)EtOAc. The yield was 0.053 g.

m/z 686/688(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ11.68 (s, 1H), 10.23 (s, 1H), 8.91 (brd, J = 26.7Hz, 2H), 8.03 (s, 1H), 7.68-7.56 ( m,1H), 7.54-7.40 (m, 2H), 6.93 (d, J = 8.5 Hz, 1H), 6.78 (d, J = 8.2 Hz, 1H), 6.56-6.43 (m, 1H), 4.91 (brd) , J=8.5Hz, 1H), 4.57-4.43 (m, 2H), 3.90-3.42 (m, 6H), 3.38-3.25 (m, 4H), 3.24-3.02 (m, 7H), 2.08 (brd, J =13.3 Hz, 2H), 1.92-1.54 (m, 2H), 1.34 (d, J = 6.7 Hz, 6H). An unobserved exchangeable proton.

Example 81

(R)-5-(2-(2-chloro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetate

a)(R)-5-(1-( third -butyldimethylmethylalkyloxy)-2-(2-chloro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)-8-hydroxyquinoline-2(1H)-one

( R )-5-(2-Amino-1-(tris-butyldimethylsilyloxy)ethyl)-8-hydroxyquinoline-2(1H)-one (WO 2004106333) (0.261 g) The solution in methanol (2 mL) was taken with EtOAc (EtOAc &lt Subsequent addition of 2-(2-chloro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 a solution of -methyl)phenyl)acetaldehyde (Example 80, step f) (0.247 g) in MeOH (3 mL), and the mixture was stirred at room temperature for 10 min then cooled in ice-water. Treated with sodium cyanoborohydride (0.102 g). The cooling bath was removed and the mixture was stirred at room temperature overnight. More sodium cyanoborohydride (0.101 g) was added and the mixture was stirred over a second night. The solution is then concentrated in vacuo on a fast cerium oxide, and the resulting powder is purified by flash chromatography on cerium oxide, eluting with 1:3:96 to 1:5:94 triethylamine:methanol:dichloromethane , a yellow oil that produces a sub-title compound. The yield was 0.130 g.

¹ H NMR (400 MHz, D _{6 -} DMSO, 90 ° C) δ 8.40 (d, J = 9.7 Hz, 1H), 8.10 (s, 1H), 7.48 (dd, J = 7.3, 2.1 Hz, 1H), 7.38- 7.30 (m, 2H), 7.17 (d, J = 8.2 Hz, 1H), 7.09 (d, J = 8.2 Hz, 1H), 6.65 (d, J = 9.7 Hz, 1H), 5.26 (dd, J = 7.6) , 5.0 Hz, 1H), 3.91-3.78 (m, 6H), 3.72 (s, 2H), 3.55-3.46 (m, 1H), 3.10-3.03 (m, 1H), 2.99 (s, 4H), 2.93 2.86 (m, 1H), 2.68-2.47 (m, 4H), 1.94-1.84 (m, 2H), 1.80-1.68 (m, 2H), 1.55 (d, J = 7.0 Hz, 6H), 0.99 (s, 9H), 0.18 (d, J = 4.6 Hz, 6H). Three unobserved exchangeable protons.

b) (R)-5-(2-(2-chloro-3-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetate

( R )-5-(1-(Third-butyldimethylmethylalkyloxy)-2-(2-chloro-3-((4-(2-isopropylthiazole-4-carbonyl)-) 1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)-8-hydroxyquinoline-2(1H)-one (Example 81, step a) (0.124 g), EtOAc (EtOAc m. The residue was dissolved in methanol (4mL), the resulting solution was filtered and purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in acetonitrile 10 to 30%) by. The product containing fractions were concentrated in vacuo and co-evaporated three times from acetonitrile to give a colourless residue. The residue was triturated with EtOAc (EtOAc)EtOAc. The yield was 0.076 g.

m/z 680/682(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.17 (d, J = 10.0 Hz, 1H), 7.94 (s, 1H), 7.55 (d, J = 7.2 Hz, 1H), 7.47-7. (m, 2H), 7.14 (d, J = 8.2 Hz, 1H), 6.99 (d, J = 8.2 Hz, 1H), 6.55 (d, J = 10.0 Hz, 1H), 5.34 (dd, J = 9.1, 4.0 Hz, 1H), 4.33-4.09 (br m, 2H), 3.77-3.60 (m, 6H), 3.44 - 2.87 (m, 11H), 2.02-1.87 (m, 2H), 1.81-1.65 (m, 2H) ), 1.35 (d, J = 6.9 Hz, 6H). Six unobserved exchangeable protons.

Example 82

(R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(5-isopropylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-(3-(1-oxa-4,9-diazaspiro[5.5]undecal-9-ylmethyl)phenyl)ethanol

Add '880' ammonia solution (5 mL) to 2,2,2-trifluoro-1-(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diaza Heterospiro[5.5]undecane-4-yl)ethanone (Example 12, step e) (2.02 g) in MeOH (25 mL). The resulting mixture was stirred for 90 min and the solvent was evaporated. The residue was purified by silica gel chromatography eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM: methanol: ' The fractions containing the product are combined and evaporated to give the clear oil of the sub-title compound. The yield was 1.44 g.

m/z 291(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.18 (t, J = 7.4 Hz, 1H), 7.13 - 7.02 (m, 3H), 4.34 (s, 1H), 3.61 (t, J = 6.9 Hz, 2H), 3.51-3.45 (m, 2H), 3.41 (s, 2H), 2.71 (t, J = 6.9 Hz, 2H), 2.62 (t, J = 4.9 Hz, 2H), 2.52 (s, 2H), 2.42 - 2.23 (m, 4H), 1.82-1.72 (m, 2H), 1.53-1.38 (m, 2H). An unobserved exchangeable proton.

b) (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(5-isopropyl Thiophen-3-yl)methanone

Adding O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.55 g) to 2-(3-(1- Oxa-4,9-diazaspiro[5.5]undecane-9-ylmethyl)phenyl)ethanol (Example 82, step a) (0.32 g), 5-isopropylthiophene-3 - a solution of formic acid (0.19 g) and triethylamine (0.61 mL) in DMF (7 mL) EtOAc. The resulting yellow solution was allowed to warm to RT and stirred for 2 h. The mixture was partitioned between EtOAc (EtOAc) (EtOAc)EtOAc. The resulting gum was purified by silica gel chromatography eluting with 47.5:47.5:5 isohexane:ethyl acetate:triethylamine to 95:5 ethyl acetate:triethylamine. To the residue containing the product, toluene (200 mL) was evaporated. The yield is 0.4g.

m/z 443(M+H) ⁺ (APCI)

c) (R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(5-isopropylthiophen-3-carbonyl)-1-oxa-4,9-diaza) Hetero[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

Adding trifluoroacetic acid (0.035 mL) to (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4- (5-isopropylthiophen-3-yl)methanone (Example 82, step b) (0.2 g) in DCM (5 mL). The mixture was stirred for 5 min, then Dess-Martin periodinane (0.29 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (5 mL) followed by acetic acid (0.026 mL) and (R)-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.14 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.043 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The title compound was obtained as a white solid. The yield was 0.07 g.

m/z 651(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ7.50-7.32 (m, 5H), 6.96-6.88 (m, 2H), 6.77 (d, J = 8.2Hz, 1H), 4.98-4.88 ( m,1H), 4.28 (s, 2H), 3.72-3.63 (m, 2H), 3.59-3.34 (m, 4H), 3.29-2.97 (m, 11H), 2.11-1.93 (m, 2H), 1.86- 1.68 (m, 2H), 1.28 (d, J = 6.7 Hz, 6H). Six unobserved exchangeable protons.

Example 83

(R)-8-hydroxy-5-(1-hydroxy-2-(3-((4-(5-isopropylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)methyl)phenethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

Add TFA (0.04 mL) to (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl) (5-Isopropylthiophen-3-yl)methanone (Example 82, step b) (0.2 g) in DCM (5 mL) The reaction was stirred for 5 min, then Dess-Martin periodinane (0.29 g) was added. The mixture was allowed to warm to RT and stirred for 1 h. Saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL). The layers were separated and the aqueous was extracted with ethyl acetate (20 mL). The combined organics were washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was redissolved in methanol (5mL), acetic acid (0.03 mL) and (R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8 -Hydroxyquinolin-2(1H)-one (WO 2004106333) (0.15 g), then the mixture was stirred for 5 min and cooled in an ice bath. Sodium cyanoborohydride (0.043 g) was then added, which was allowed to warm to RT and stirred overnight. The reaction was concentrated and purified by silica gel chromatography eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM: methanol: &lt; The fractions containing the product are combined and evaporated. The residue was dissolved in THF (5 mL) EtOAc (EtOAc) The solvent was evaporated and the residue was azeotroped twice with toluene. Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 10 to 30% acetonitrile). The title compound was obtained as a white solid. The yield was 0.06 g.

m/z 645(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.17 (d, J = 9.7 Hz, 1H), 7.48 (s, 1H), 7.45-7.33 (m, 4H), 7.14 (d, J = 8.2 Hz, 1H), 7.00 (d, J = 8.2 Hz, 1H), 6.90 (s, 1H), 6.54 (d, J = 10.0 Hz, 1H), 5.36 (dd, J = 8.5, 4.1 Hz, 1H) , 4.29 (s, 2H), 3.72-3.63 (m, 2H), 3.57-3.38 (m, 4H), 3.28 (t, J = 8.1 Hz, 2H), 3.22-2.98 (m, 9H), 2.10. (m, 2H), 1.81-1.62 (m, 2H), 1.28 (d, J = 6.7 Hz, 6H). Six unobserved exchangeable protons.

Example 84

(R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(2-isobutylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-isobutyl Thiazol-4-yl)methanone

Adding O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.34 g) to 2-(3-(1- Oxa-4,9-diazaspiro[5.5]undecane-9-ylmethyl)phenyl)ethanol (Example 82, step a) (0.2 g), 2-isobutylthiazole-4 - a solution of formic acid (0.13 g) and triethylamine (0.38 mL) in DMF (7 mL) EtOAc. The resulting yellow solution was allowed to warm to RT and stirred for 2 h. The mixture was partitioned between EtOAc (EtOAc) (EtOAc)EtOAc. The resulting gum was purified by silica gel chromatography eluting with 47.5:47.5:5 isohexane:ethyl acetate:triethylamine to 95:5 ethyl acetate:triethylamine. To the residue containing the product, toluene (200 mL) was evaporated. The yield was 0.2 g.

m/z 458(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D6-DMSO, 90 ° C) δ 7.79 (s, 1H), 7.26-7.03 (m, 4H), 4.60 (t, J = 5.3 Hz, 1H), 3.72-3.50 (m, 8H) ), 3.46-3.34 (m, 2H), 2.94-2.84 (m, 2H), 2.71 (t, J = 7.1 Hz, 2H), 2.43-2.01 (m, 5H), 1.74-1.37 (m, 4H), 1.01-0.90 (m, 6H)

b) (R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(2-isobutylthiazole-4-carbonyl)-1-oxa-4,9-diaza) Hetero[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

Trifluoroacetic acid (0.032 mL) was added to (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[Delta] at 0 °C in DCM (5 mL). 5.5]undecane-4-yl)(2-isobutylthiazol-4-yl)methanone (Example 84, step a) (0.19 g). The mixture was stirred for 5 min, then Dess-Martin periodinane (0.26 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (5 mL) followed by acetic acid (0.024 mL) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.11 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.04 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 35% acetonitrile). The title compound was obtained as a white solid. Yield 0.Ig.

m/z 666(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.27 (s, 1H), 7.93 (s, 1H), 7.44-7.33 (m, 4H), 6.93 (d, J = 8.5Hz, 1H) , 6.77 (d, J = 8.2 Hz, 1H), 4.92 (dd, J = 7.9, 5.1 Hz, 1H), 4.34 - 4.21 (m, 2H), 3.73 - 3.60 (m, 6H), 3.28-2.96 (m , 10H), 2.88 (d, J = 6.9 Hz, 2H), 2.11-1.95 (m, 3H), 1.84-1.64 (m, 2H), 0.95 (d, J = 6.7 Hz, 6H). Five unobserved exchangeable protons.

Example 85

(R)-7-(2-(2-Fluoro-3-((4-(6-isopropylpyridine)methyl)-1-oxa-4,9-diazaspiro[5.5] Cycloalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 2-isopropylpyridine

A solution of butyllithium (1.6 M in hexanes, 50 mL) was added dropwise to a solution of 2-ethylpyridine (9.34 mL) in tetrahydrofuran (50 mL) in EtOAc. The mixture was stirred for 2 h, then methyl iodide (5 mL) was added dropwise over 15 min. The resulting orange slurry was allowed to warm to RT and stirred overnight. The solvent was evaporated and the residue diluted with diethyl ether (100 mL). The organic layer was washed with EtOAc (EtOAc)EtOAc. The yield was 9.1 g.

m/z 122 (M+H) ⁺ (APCI) ¹ H NMR (300MHz, CDCl ₃ ) δ 8.56-8.52 (m, 1H), 7.60 (td, J = 7.7, 2.1 Hz, 1H), 7.19-7.15 (m, 1H), 7.09 (ddd, J = 7.4, 4.9, 1.2 Hz, 1H), 3.06 (seven peak, J = 6.9 Hz, 1H), 1.31 (d, J = 6.9 Hz, 6H).

b) 2-isopropylpyridine 1-oxide

MCPBA (22.0 g) was added to a solution of 2-isopropylpyridine (Example 85, Step a) (9.1 g) in DCM (250 mL) The reaction mixture was washed with aq. EtOAc (EtOAc) Purified by silica gel chromatography eluting EtOAc (EtOAc) elut The fractions containing the product are combined and evaporated to give the sub-title compound as a yellow oil. The yield was 3.9 g.

m/z 138(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO) δ 8.23 (dd, J = 6.2, 1.5 Hz, 1H), 7.41 (dd, J = 7.7, 2.3 Hz, 1H), 7.35-7.24 (m, 2H), 3.56 (seven peak, J=7 Hz, 1H), 1.20 (d, J=7 Hz, 6H).

c) 6-isopropylpyridine carbonitrile

Trimethyldecyl cyanide (1.53 mL) was added to a solution of 2-isopropylpyridine 1-oxide (Example 85, step b) (1.3 g) in DCM (40 mL). . Diethylaminoformamidine chloride (1.2 mL) was added and the mixture was stirred for 3 days. Aqueous potassium carbonate (10%, 40 mL) was added and the mixture was stirred for 10 min. The layers were separated and the aqueous extracted with DCM (2×40 mL). The combined organic solution was washed with brine (40 mL) dried over sodium sulfate. Purified by silica gel chromatography and eluted with 3:1 isohexane:ethyl acetate. The fractions containing the product are combined and evaporated to give the clear oil of the sub-title compound. The yield was 1.23 g.

m/z 147(M+H) ⁺ (APCI)

_{^{1 H NMR (300MHz, CDCl 3}} ) δ7.74 (t, J = 7.8Hz, 1H), 7.51 (dd, J = 7.6,0.9Hz, 1H), 7.39 (dd, J = 8.1,0.9Hz, 1H) , 3.11 (seven peaks, J = 6.7 Hz, 1H), 1.31 (d, J = 6.7 Hz, 6H).

d) 6-isopropyl picolinic acid

Concentrated hydrochloric acid (15 mL) was added to a solution of 6-isopropylpyridinecarbonitrile (Example 85, step c) (1.23 g) in methanol (30 mL). The resulting mixture was heated under reflux for 17 h and allowed to cool to RT. The mixture was cautiously poured into a sodium hydroxide solution (10M, 50 mL) and stirred at RT overnight. The reaction was concentrated and the pH was adjusted to 5 using 2M HCl solution. The aqueous mixture was extracted with chloroform (3×100 mL). The organic solution was combined with EtOAc (EtOAc m. The yield was 0.94 g.

m/z 166(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO) δ 7.89-7.81 (m, 2H), 7.48 (dd, J = 7.2, 1.3 Hz, 1H), 3.05 (seven peaks, J = 6.9 Hz, 1H), 1.22 (d, J = 6.9 Hz, 6H). An unobserved exchangeable proton.

e) (9-(2-Fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(6 -isopropylpyridin-2-yl)methanone

Adding O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.20 g) to 9-(3-(2- ( Third -butyldimethylsilyloxy)ethyl)-2-fluorobenzyl)-1-oxa-4,9-diazaspiro[5.5]undecane (Example 78, Step d) (0.17 g), 6-isopropylpicolinic acid (Example 85, step d) (0.07 g) and triethylamine (0.22 mL) in DMF (7 mL) in EtOAc. The resulting yellow solution was allowed to warm to RT and stirred for 2 h. The mixture was partitioned between ethyl acetate (100 mL) and brine (100 mL). The organic phase was separated, washed with brine (2×100 mL) dried over sodium sulfate. The residue was dissolved in THF (10 mL) EtOAc (EtOAc) The resulting mixture was stirred for 2 h and the solvent was evaporated. Purified by silica gel chromatography, eluting with 47.5:47.5:5 isohexane:ethyl acetate:triethylamine to 95:5 ethyl acetate:triethylamine gradient. To the residue containing the product, toluene (50 mL) was evaporated. The yield was 0.2 g.

m/z 456(M+H) ⁺ (APCI)

f) (R)-7-(2-(2-fluoro-3-((4-(6-isopropylpyridine))-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

Add TFA (0.034 mL) to (9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 4-Base)(6-isopropylpyridyl-2-yl)methanone (Example 85, step e) (0.2 g) in DCM (5 mL) The mixture was stirred for 5 min, then Dess-Martin periodinane (0.28 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (5 mL) followed by acetic acid (0.025 mL) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.12 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.04 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 35% acetonitrile). The title compound was obtained as a white solid. The yield was 0.057 g.

m/z 664(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.27 (s, 1H), 7.81 (t, J = 7.7 Hz, 1H), 7.54-7.21 (m, 5H), 6.94 (d, J = 8.2 Hz, 1H), 6.77 (d, J = 8.5 Hz, 1H), 4.97-4.82 (m, 1H), 4.40-4.25 (m, 2H), 3.77-3.47 (m, 6H), 3.29-2.99 (m) , 11H), 2.11 - 1.62 (m, 4H), 1.24 (d, J = 6.9 Hz, 6H). Five unobserved exchangeable protons.

Example 86

(R)-7-(2-(3-((4-(5-ethylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (5-ethylthiophen-3-yl)(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane -4-yl)methanone

Add HATU (0.209 g) to 5-ethylthiophene-3-carboxylic acid (0.086 g), 2-(3-(1-oxa-4,9-diazaspiro[5.5]undecane-9 a stirred solution of -methyl)phenyl)ethanol (Example 82, Step a) (0.160 g) and triethylamine (0.3 mL) in DMF (2 mL). After 1 h the reaction mixture was diluted with EtOAc (EtOAc) The ethyl acetate layer was evaporated in vacuo. Purification by silica gel chromatography eluting with 10:1 ethyl acetate: triethylamine afforded the subtitle compound. The yield was 0.18 g.

¹ H NMR (400 MHz, D6-DMSO, 90 ° C) δ 7.52 (s, 1H), 7.20 (t, J = 7.4 Hz, 1H), 7.13 - 7.05 (m, 3H), 6.90 (s, 1H), 4.60 (t, J = 5.3 Hz, 1H), 3.64 - 3.55 (m, 4H), 3.53-3.46 (m, 2H), 3.31 (s, 4H), 2.81 (q, J = 7.3 Hz, 2H), 2.74 -2.66 (m, 2H), 2.37-2.23 (m, 4H), 1.75-1.65 (m, 2H), 1.56-1.35 (m, 2H), 1.24 (t, J = 7.7 Hz, 3H).

b) 2-(3-((4-(5-ethylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl Phenyl) acetaldehyde

Add Dess-Martin periodinane (0.232 g) to (5-ethylthiophen-3-yl)(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9 - a stirred solution of diazaspiro[5.5]undec-4-yl)methanone (Example 86, Step a) (0.180 g) and trifluoroacetic acid (0.042 mL) in DCM (5 mL). After 1 h, ethyl acetate (30 mL) was added followed by a mixture of saturated sodium thiosulfate (5 mL) and saturated sodium bicarbonate (5 mL). The reaction mixture was thoroughly shaken and separated. The ethyl acetate solution was washed with saturated sodium bicarbonate solution, water and brine. Acetic acid (0.08 mL) was added, the solution was dried over sodium sulfate, filtered and evaporated in vacuo. The yield was 0.17 g. Use directly.

m/z 427(M+H) ⁺ (APCI)

c) (R)-7-(2-(3-((4-(5-ethylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane -9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

Acetic acid (0.036 mL) was added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.166 g) and 2-(3-((4-(5-ethylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5] A solution of the monoalkane-9-yl)methyl)phenyl)acetaldehyde (Example 86, step b) (0.180 g) in methanol (EtOAc) After 1 min, sodium cyanoborohydride (0.080 g) was added. 1.5h to, reaction mixture was filtered and purified by preparative HPLC (Sunfire ^TM, Gradient by: in aqueous 0.2% TFA in 10 to 40% acetonitrile binding solution containing the pure product of the fractions were evaporated in vacuo Acetonitrile (200mL) and. The solution was evaporated in vacuo to give a crystallite.

m/z 637(M+H) ⁺ (APCI)

¹ H NMR (300 MHz, D ₆ -DMSO, 90 ° C) δ 11.37 (s, 1H), 7.51-7.30 (m, 5H), 6.93 (d, J = 8.1 Hz, 1H), 6.77 (d, J = 8.1 Hz, 1H), 6.91-6.86 (m, 1H), 4.94-4.86 (m, 1H), 4.29 (s, 2H), 3.71-3.31 (m, 8H), 3.29-2.93 (m, 8H), 2.81 (q, J = 7.9 Hz, 2H), 2.15 - 1.91 (m, 2H), 1.75-1.51 (m, 2H), 1.25 (t, J = 7.6 Hz, 3H). Five unobserved exchangeable protons.

Example 87

( R -4-hydroxy-7-(1-hydroxy-2-(3-((4-(2-propylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) butane thioxamide

Phosphorus pentasulfide (3 g) was added to a suspension of butaamine (5 g) in MTBE (300 mL) and the mixture was stirred for 3 h. The reaction was filtered through celite and the pad was washed with EtOAc (EtOAc). The combined filtrate and washings were evaporated to give the subtitle compound as a yellow oil. The yield was 5.2 g.

_{^{1 H NMR (300MHz, D 6}} -DMSO) δ9.32 (s, 1H), 9.12 (s, 1H), 2.43 (t, J = 7.4Hz, 2H), 1.72-1.59 (m, 2H), 0.86 ( t, J = 7.4 Hz, 3H).

b) Ethyl 2-propylthiazole-4-carboxylate

Add ethyl 3-bromo-2-oxopropionate (6.32 mL) to a solution of butane thioguanamine (Example 87, step a) (5.2 g) in ethanol (100 mL) and a mixture Heat overnight under reflux. The solvent was evaporated and the residue was crystalljjjjjjjjjjjj The layers were separated and the aqueous extracted with ethyl acetate (2×100 mL). The combined organic solution was washed with brine (100 mL) dried over sodium sulfate. Purified by silica gel chromatography and eluted with 10:1 isohexane:ethyl acetate. The fractions containing the product are combined and evaporated to give the sub-title compound as a yellow oil. The yield was 5.24 g.

m/z 200(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.05 (s, 1H), 4.42 (q, J = 7.2 Hz, 2H), 3.04 (t, J = 7.7 Hz, 2H), 1.89-1.78 (m, 2H) , 1.40 (t, J = 7.2 Hz, 3H), 1.02 (t, J = 7.2 Hz, 3H).

c) 2-propylthiazole-4-carboxylic acid

To a mixture of THF (80 mL) and water (20 mL) In solution. The resulting mixture was stirred overnight. The reaction was acidified with concentrated hydrochloric acid and the volatiles were evaporated. The aqueous mixture formed was saturated with sodium chloride and extracted with ethyl acetate (3×100 mL). The combined organic solution was dried with sodium sulfate, filtered and evaporated elut The yield is 2.5g.

_{^{1 H NMR (300MHz, D 6}} -DMSO) δ 12.91 (s, 1H), 8.31 (s, 1H), 2.97 (t, J = 7.5Hz, 2H), 1.81-1.67 (m, 2H), 0.95 (t , J = 7.3 Hz, 3H).

d) (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-propylthiazole -4-yl)methanone

Adding O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.24 g) to 2-(3-(1- Oxa-4,9-diazaspiro[5.5]undecane-9-ylmethyl)phenyl)ethanol (Example 82, step a) (0.14 g), 2-propylthiazole-4- Formic acid (Example 87, step c) (0.084 g) and triethylamine (0.27 mL) in EtOAc (EtOAc) The resulting yellow solution was allowed to warm to RT and stirred for 2 h. The mixture was partitioned between EtOAc (EtOAc) (EtOAc)EtOAc. The resulting gum was purified by silica gel chromatography eluting with 47.5:47.5:5 isohexane:ethyl acetate:triethylamine to 95:5 ethyl acetate:triethylamine. To the residue containing the product, toluene (200 mL) was evaporated. The yield was 0.14 g.

m/z 444(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.91 (s, 1H), 7.18 (t, J = 7.4 Hz, 1H), 7.12 - 7.04 (m, 3H), 4.37 - 4.30 (m, 1H), 3.69-3.57 (m, 8H), 3.41 (s, 2H), 2.98 (t, J = 7.3 Hz, 2H), 2.71 (t, J = 7.1 Hz, 2H), 2.40-2.23 (m, 4H) ), 1.82-1.64 (m, 4H), 1.59-1.46 (m, 2H), 0.97 (t, J = 7.4 Hz, 3H).

e) (R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(2-propylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

Add TFA (0.023 mL) to (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl) (2-propylthiazol-4-yl)methanone (Example 87, step d) (0.13 g) in DCM (5 mL) The mixture was stirred for 5 min, then Dess-Martin periodinane (0.19 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (5mL), followed by addition of acetic acid (0.017 mL) and (R) -7- (2- amino-1-hydroxyethyl) -4-hydroxybenzo [d] thiazol -2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.077 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.028 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 35% acetonitrile). The title compound was obtained as a white solid. The yield was 0.09 g.

m/z 652(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.29 (s, 1H), 7.96-7.92 (m, 1H), 7.46-7.32 (m, 4H), 6.98-6.92 (m, 1H), 6.81-6.75 (m, 1H), 4.98-4.90 (m, 1H), 4.34-4.27 (m, 2H), 3.76-3.62 (m, 6H), 3.30-2.94 (m, 12H), 2.08-1.98 (m) , 2H), 1.85-1.70 (m, 4H), 1.01-0.92 (m, 3H). Five unobserved exchangeable protons.

Example 88

(R)-7-(2-(3-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenyl)propylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoro Acetate

a) 3-(2-fluorophenyl)propan-1-ol

The borane dimethyl sulfide solution complex (2M in THF, 27.6 mL) was added dropwise to a solution of 3-(2-fluorophenyl)propionic acid (3.09 g) in tetrahydrofuran (25 mL). The resulting mixture was warmed to RT and stirred overnight. The reaction was stopped with methanol and evaporated when bubbling ceased. Purified by silica gel chromatography and eluted with a gradient of 4:1 to 1:1 isohexane:ethyl acetate. The fractions containing the product are combined and evaporated in vacuo to give the sub-title compound as a clear oil. The yield was 2.72 g.

_{^{1 H NMR (300MHz, CDCl 3}} ) δ7.24-7.13 (m, 2H), 7.09-6.97 (m, 2H), 3.67 (t, J = 6.3Hz, 2H), 2.74 (t, J = 7.6Hz, 2H), 1.95-1.83 (m, 2H). An unobserved exchangeable proton.

b) tert-butyl(3-(2-fluorophenyl)propoxy)dimethyl decane

Third - butyldimethylsilyl silicon alkyl chloride (3.19 g of) was added to imidazole (3.6 g of) and 3- (2-fluorophenyl) propan-1-ol (Example 88, step a) (2.72g) It was cooled in an ice bath in anhydrous DMF (30 mL). After 45 min the reaction was diluted with EtOAc (EtOAc) The formed gel was purified by silica gel chromatography and dissolved in isohexane. The fractions containing the product are combined and evaporated to give the clear oil of the sub-title compound. The yield was 4.4 g.

_{^{1 H NMR (300MHz, CDCl 3}} ) δ7.23-7.09 (m, 2H), 7.09-6.94 (m, 2H), 3.64 (t, J = 6.3Hz, 2H), 2.75-2.66 (m, 2H), 1.89-1.76 (m, 2H), 0.91 (s, 9H), 0.05 (s, 6H).

c) 3-(3-( third -butyldimethylmethylalkyloxy)propyl)-2-fluorobenzaldehyde

Tri -butyl(3-(2-fluorophenyl)propoxy)dimethyloxane (Example 88, step b) (4.4 g) was added dropwise to the second butyllithium in 5 min (in cyclohexane) 1.4 M in alkane, 11.7 mL) and 1,1,4,7,7-pentamethyldiethylethylamine (3.4 mL) in THF (25 mL). The resulting mixture was stirred for 2 h then DMF (EtOAc &lt The reaction was quenched with water (100 mL) and then ethyl acetate (250 mL). The phases were separated and the organic phase was washed with water (2 <RTI ID=0.0></RTI></RTI><RTIgt; Purification by gel chromatography eluting with a gradient of 10% diethyl ether from isohexane to isohexane. The product-containing fractions are combined and evaporated to give the sub-title compound as a clear oil. The yield is 1g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ10.38 (s, 1H), 7.73-7.67 (m, 1H), 7.48 (td, J = 7.4,1.8Hz, 1H), 7.18 (t, J = 7.7Hz, 1H), 3.66 (t, J = 6.0 Hz, 2H), 2.82-2.75 (m, 2H), 1.90-1.80 (m, 2H), 0.91 (s, 9H), 0.06 (s, 6H).

d) (9-(2-Fluoro-3-(3-hydroxypropyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2 -isopropylthiazol-4-yl)methanone

Add 3-(3-( Third -butyldimethylsulfonyloxy)propyl)-2-fluorobenzaldehyde (Example 88, step c) (0.15 g) to (2-isopropylthiazole) 4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (0.19 g) and Acetic acid (0.03 mL) in a solution of N-methyl-2-pyrrolidone (10 mL). The resulting mixture was stirred for 15 min and then cooled in an ice bath. Sodium triethoxy borohydride (0.16 g) was then added and the mixture was stirred overnight. The reaction was poured into a mixture of saturated sodium bicarbonate (20 mL) and water (100 mL). The aqueous solution was extracted with ethyl acetate (3×100 mL). The combined organics were washed with water (50 mL) brine brine The residue was redissolved in THF (10 mL) and EtOAc (EtOAc m. The resulting mixture was stirred for 2 h and the solvent was evaporated. The residue was purified by column chromatography eluting with 4:1 EtOAc:EtOAcEtOAcEtOAc The fractions containing the product are combined and evaporated to give the clear oil of the sub-title compound. The yield was 0.22 g.

m/z 476(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D6-DMSO, 90 ° C) δ 7.99 (s, 1H), 7.24 - 7.12 (m, 2H), 7.08 - 7.01 (m, 1H), 4.48 (t, J = 5.1 Hz, 1H) ), 3.74-3.38 (m, 10H), 2.62 (t, J = 7.7 Hz, 2H), 2.46-2.13 (m, 4H), 1.74-1.44 (m, 6H), 1.35 (d, J = 6.4 Hz, 6H). A proton that is blurred by the water peaks.

e) (R)-7-(2-(3-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)methyl)phenyl)propylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di- Trifluoroacetate

Adding trifluoroacetic acid (0.032 mL) to (9-(2-fluoro-3-(3-hydroxypropyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undec carbon Alkyl-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 88, step d) (0.2 g) in DCM (5 mL) The mixture was stirred for 5 min, then Dess-Martin periodinane (0.27 g) was added. The resulting yellow solution was allowed to warm to RT and stirred for 1 h. A mixture of saturated sodium thiosulfate solution (5 mL), sat. sodium bicarbonate (5 mL) and ethyl acetate (20 mL) was then added and the mixture was stirred vigorously for 10 min. The aqueous phase was separated and extracted with ethyl acetate (20 mL). The combined organic solution was washed with brine (20 mL) EtOAc (EtOAc)EtOAc. The residue was dissolved in methanol (5 mL) followed by acetic acid (0.024 mL) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2 (3H) - Ketone hydrochloride (WO 2007027134, Example 1, step d) (0.11 g), and the mixture was stirred for 5 min then cooled in an ice bath. Sodium cyanoborohydride (0.040 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 35% acetonitrile). The title compound was obtained as a white solid. The yield was 0.14 g.

m/z 684(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.25 (s, 1H), 7.98-7.89 (m, 1H), 7.52-7.35 (m, 2H), 7.28-7.16 (m, 1H), 6.98-6.86(m,1H), 6.82-6.70(m,1H),4.96-4.84(m,1H), 4.39-4.24(m,2H),3.77-3.58(m,6H),3.34-2.94(m , 9H), 2.80-2.66 (m, 2H), 2.10 - 1.70 (m, 6H), 1.41-1.27 (m, 6H) and 5 unobserved exchangeables

Example 89

(R)-4-hydroxy-7-(1-hydroxy-2-(3-((10-(2-methylthiazole-4-carbonyl)-7-oxa-3,10-diazaspiro[ 5.6] Dodecane-3-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) tert-butyl 4-(cyanomethyl)-4-hydroxypiperidine-1-carboxylate

A solution of 1-oxa-6-azaspiro[2.5]octane-6-carboxylic acid tert-butyl ester (2 g) in DMF (20 mL). Stir for 4 days. The mixture was partitioned between EtOAc and EtOAc. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 1.18 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ3.95-3.85 (m, 2H), 3.19-3.10 (m, 2H), 2.54 (s, 2H), 1.86 (s, 1H), 1.76-1.61 (m, 4H ), 1.46 (s, 9H).

b) tert-butyl 4-(2-aminoethyl)-4-hydroxypiperidine-1-carboxylate

a solution of 4-(cyanomethyl)-4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (Example 89, step a) (1.4 g) in a mixture of ethanol (20 mL) and acetic acid (20 mL) Hydrogenation was carried out in the presence of platinum (IV) oxide (0.25 g) at 4 atmospheres of hydrogen for 4 hours. The catalyst was filtered off and the solvent was removed under reduced pressure. The residue was partitioned between EtOAc EtOAc m. The yield is 1.1g. Use directly.

c) 4-(2-(2-chloroethylguanidino)ethyl)-4-hydroxypiperidine-1-carboxylic acid tert-butyl ester

Chloroacetyl chloride (0.483 mL) was added dropwise to 10-butanyl 4-(2-aminoethyl)-4-hydroxypiperidine-1-carboxylate in ethyl acetate (25 mL) over 10 min (Example 89 Step b) (1.1 g) and potassium carbonate (1.77 g) dissolved in water (20 mL) were stirred vigorously at 0 °C. The mixture was then stirred at 0 ° C for 45 minutes and then extracted with ethyl acetate. The organic layer was dried with sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.9 g.

m/z 319(MH) ^- (APCI)

d) 9-Sideoxy-7-oxa-3,10-diazaspiro[5.6]dodecane-3-carboxylic acid tert-butyl ester

3-(2-(2-Chloroethylamino)ethyl)-4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (Example 89, step c) (0.3 g) in dry THF (18 mL) The solution was added dropwise to a refluxing mixture of potassium succinate (1M in EtOAc) (3 mL) and dry THF (60 mL). At the end of the addition, the mixture was heated under reflux for another 15 minutes and then cooled to room temperature. The mixture was partitioned between ethyl acetate and brine. The organic layer was dried with sodium sulfate, filtered and evaporated. The sub-title compound was obtained by wet milling with diethyl ether. The yield was 0.117 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ 7.67 (s, 1H), 4.01 (s, 2H), 3.57 (d, J = 13.1Hz, 2H), 3.11 (dd, J = 9.6,4.2Hz, 2H ), 3.05-2.95 (m, 2H), 1.85-1.76 (m, 4H), 1.38 (s, 9H), 1.37-1.32 (m, 2H).

e) 7-oxa-3,10-diazaspiro[5.6]dodecane-3-carboxylic acid tert-butyl ester

Add borane-methylthio complex (2M in THF, 2.88 mL) to 9-oxooxy-7-oxa-3,10-diazaspiro[5.6]dodecane-3- A solution of the third butyl formate (Example 89, step d) (0.41 g) in dry THF (40 mL) was then evaporated. The mixture was cooled to room temperature and quenched with methanol. The solvent was removed under reduced pressure and the residue was evaporated mjjjjjjj N1, N2-dimethylethane-1,2-diamine (1.0 g) was added, and the mixture was heated under nitrogen under nitrogen for 16 h. Additional N1 , N2 -dimethylethane-1,2-diamine (1.0 g) was added and reflux was continued for 16 h. The solvent was evaporated under reduced pressure and the residue was azeotroped with toluene. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.176 g.

m/z 271(M+H) ⁺ (APCI)

f) 10-(2-methylthiazole-4-carbonyl)-7-oxa-3,10-diazaspiro[5.6]dodecane-3-carboxylic acid tert-butyl ester

HATU (0.322 g) was added in one portion to 7-oxa-3,10-diazaspiro[5.6]dodecane-3-carboxylic acid tert-butyl ester (Example 89, step e) (0.176 g) and 2 Methylthiazole-4-carboxylic acid (0.093 g) and triethylamine (0.36 mL) in EtOAc (10 mL) EtOAc. The mixture was then stirred at 20 ° C for 1 hour. The reaction mixture was partitioned between ethyl acetate and brine. The organic layer was washed twice with brine, dried over sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 210 mg.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 7.78 (s, 1H), 3.69 (s, 4H), 3.65-3.62 (m, 2H), 3.59-3.53 (m, 2H), 3.11 3.03 (m, 2H), 2.66 (s, 3H), 1.85-1.81 (m, 2H), 1.74-1.67 (m, 2H), 1.39 (s, 9H), 1.38-1.33 (m, 2H).

g) (2-methylthiazol-4-yl)(7-oxa-3,10-diazaspiro[5.6]dodecan-10-yl)methanone trifluoroacetate

3-(2-methylthiazole-4-carbonyl)-7-oxa-3,10-diazaspiro[5.6]dodecane-3-carboxylic acid tert-butyl ester (Example 89, step f) The solution (0.21 g) in DCM (10 mL) Toluene (40 mL) was added and the solvent was evaporated under reduced pressure. The residue is azeotroped with acetonitrile to give the sub-title compound. The yield was 0.21 g.

m/z 296(M+H) ⁺ (APCI)

h) (3-(3-(2-hydroxyethyl)benzyl)-7-oxa-3,10-diazaspiro[5.6]dodecane-10-yl)(2-methylthiazole -4-yl)methanone

(2-methylthiazol-4-yl)(7-oxa-3,10-diazaspiro[5.6]dodecan-10-yl)methanone trifluoroacetate (Example 89, step g (0.21 g) in acetonitrile (15 mL) was treated with triethylamine (0.214 mL) followed by 2-(3-(bromomethyl)phenyl)ethanol (Example 6, step a) (0.121 g) )deal with. The reaction mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure and the residue was partitioned between DCM and sat. The aqueous layer was re-extracted twice with DCM and the combined organics dried over sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.21 g.

m/z 430(M+H) ⁺ (APCI)

i)( R -4-hydroxy-7-(1-hydroxy-2-(3-((10-(2-methylthiazole-4-carbonyl)-7-oxa-3,10-diazaspiro[5.6] Dodecane-3-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

(3-(3-(2-hydroxyethyl)benzyl)-7-oxa-3,10-diazaspiro[5.6]dodecan-10-yl)(2-methylthiazole-4 - ketone (Example 89, step h) (0.21 g) in EtOAc (EtOAc:EtOAc. The resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.028 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.193 g) and acetic acid (0.028 mL) in methanol (15 mL). The mixture was cooled in an ice-bath and treated with sodium <RTI ID=0.0> The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.145 g.

m/z 638(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.27 (s, 1H), 7.79 (s, 1H), 7.44 - 7.32 (m, 4H), 6.93 (d, J = 8.5 Hz, 1H) , 6.77 (d, J = 8.5 Hz, 1H), 4.94 - 4.89 (m, 1H), 4.28 (s, 2H), 3.71 (s, 4H), 3.68-3.64 (m, 2H), 3.25 (t, J = 8.1 Hz, 2H), 3.20-2.99 (m, 8H), 2.66 (s, 3H), 2.04-1.95 (m, 2H), 1.91-1.83 (m, 2H), 1.78-1.67 (m, 2H). Five unobserved exchangeable protons.

Example 90

(R)-7-(2-(2-(2-isopropyl-3-(yl)-3-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenyl)-2-methylpropylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)- Keto di-trifluoroacetate

a) methyl 2-(2-fluorophenyl)acetate

A solution of 2-(2-fluorophenyl)acetic acid (9.6 g) in MeOH (EtOAc) (EtOAc) The solvent was removed under reduced pressure and the residue was crystalljjjjjjjjjjjjjj The yield was 9.7 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.29-7.23 (m, 2H), 7.13 - 7.03 (m, 2H), 3.71 (s, 3H), 3.68 (s, 2H).

b) Methyl 2-(2-fluorophenyl)-2-methylpropanoate

A solution of methyl iodide (3.2 mL) in dry DMF (80 mL) EtOAc (EtOAc &lt;RTI ID=0.0&gt; a) (2.75g). The mixture was allowed to warm slowly to room temperature and then stirred at room temperature for 5 hours. The reaction was quenched by the careful addition of saturated aqueous ammonium chloride (120 mL). The mixture was extracted with EtOAc (EtOAc) EtOAc. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 2.6 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ7.46-7.40 (m, 1H), 7.35-7.29 (m, 1H), 7.22-7.11 (m, 2H), 3.58 (s, 3H), 1.48 (s , 6H).

c) 2-(2-fluorophenyl)-2-methylpropionic acid

Add a solution of sodium hydroxide (1 g) in water (50 mL) to methyl 2-(2-fluorophenyl)-2-methylpropanoate (Example 90, step b) (2.6 g) 50 mL) and THF (50 mL) in solution. The reaction mixture was heated at 40 ° C for 40 hours. The organics were removed under reduced pressure and the remaining aqueous was washed with ethyl acetate. The aqueous layer was cooled and acidified by the addition of concentrated HCl. The mixture was extracted with EtOAc and EtOAc (EtOAc)EtOAc. The yield is 1.5g.

m/z 181(M-H)-(APCI)

d) 2-(2-fluorophenyl)-2-methylpropan-1-ol

Borane-methylsulfide (2M in THF, 12.35 mL) was added dropwise to 2-(2-fluorophenyl)-2-methylpropanoic acid (Example 90, step c) (1.5 g) in THF In a solution (30 mL). The reaction mixture was stirred at 20 ° C for 8 hours. The mixture was quenched by the careful addition of methanol until the evolution of the gas ceased. The solvent was removed under reduced pressure and the residue was azeotroped with toluene. The crude product was purified by flash chromatography on silica gel eluting with 30% ethyl acetate in isohexane. The pure fractions were evaporated to dryness to give the subtitle compound. The yield is 1.3g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.36-7.31 (m, 1H), 7.25-7.19 (m, 1H), 7.13-7.08 (m, 1H), 7.04-6.99 (m, 1H), 3.79 (dd , J = 6.4, 1.0 Hz, 2H), 1.39 (d, J = 1.0 Hz, 6H). An unobserved exchangeable proton.

e ) third -butyl (2-(2-fluorophenyl)-2-methylpropoxy)dimethyl decane

Tri -butyldimethylbenzyl chloride (1.4 g) was added portionwise to 2-(2-fluorophenyl)-2-methylpropan-1-ol (Example 90, step d) (1.3 g) And imidazole (0.631 g) in DMF (7 mL) in a stirred solution at 20 °C. The reaction mixture was stirred at room temperature for 3 hours and then partitioned between ethyl acetate and brine. The organic layer was washed twice with brine, dried over sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on EtOAc. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 1.4g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.34-7.29 (m, 1H), 7.20-7.14 (m, 1H), 7.08-7.03 (m, 1H), 7.00-6.94 (m, 1H), 3.71 (s , 2H), 1.35 (s, 6H), 0.80 (s, 9H), -0.06 (s, 6H).

f ) 3-(1-( third -butyldimethylmethylalkyloxy)-2-methylpropan-2-yl)-2-fluorobenzaldehyde

The second butyl lithium (1.4 M in cyclohexane, 3.54 mL) was added to dry THF (10 mL) and the solution was cooled to -78. Slowly added dropwise N1- (2- (dimethylamino) ethyl) - N1, N2, N2 - trimethyl-1,2-diamine (0.859g). Then added dropwise over 5 minutes to a third - butyl (2- (2-fluorophenyl) -2-methyl-propoxy) dimethyl Silane (Example 90, step e) (1.4g) in dry tetrahydrofuran Solution in (3 mL). The reaction mixture was stirred at -78 °C for 2 hours. DMF (2.69 mL) was added dropwise over 5 min and the mixture was stirred at -78 °C for one hour and then at room temperature for 45 min. The reaction mixture was quenched by the addition of water. The acid ethyl ester (200 mL) was added and the organics were washed three times with water, then twice with 2M HCl and twice with water, then brine, and dried over sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on EtOAc. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.98 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ10.39 (s, 1H), 7.75-7.70 (m, 1H), 7.63-7.58 (m, 1H), 7.19 (t, J = 7.8Hz, 1H), 3.74 ( s, 2H), 1.39 (d, J = 1.3 Hz, 6H), 0.79 (s, 9H), -0.05 (s, 6H).

g) (9-(2-Fluoro-3-(1-hydroxy-2-methylpropan-2-yl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecene Alkyl-4-yl)(2-isopropylthiazol-4-yl)methanone

Add sodium triethoxysilane borohydride (0.502 g) to (2-isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4 -yl)methanone trifluoroacetate (Example 22, step b) (0.668 g) and 3-(1-(tris-butyldimethylsilyloxy)-2-methylpropan-2- 2-fluorobenzaldehyde (Example 90, step f) (0.49 g) and acetic acid (0.090 mL) in NMP (20 mL). The reaction mixture was then stirred at 20 ° C for 18 hours. The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The resulting gum was dissolved in THF (20 mL) eluting with EtOAc (EtOAc:EtOAc The solution was allowed to stand at 20 ° C for 8 hours. The solvent was evaporated under reduced pressure and the crude material was purified eluting eluting eluting eluting The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.63 g.

m/z 490(M+H) ⁺ (APCI)

h) (R)-7-(2-(2-(2-isopropyl-3-(thiazolidine-4-carbonyl)-1-oxa-4,9-diaza) Spiro[5.5]undecane-9-yl)methyl)phenyl)-2-methylpropylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2 (3H )-ketodi-trifluoroacetate

(9-(2-Fluoro-3-(1-hydroxy-2-methylpropan-2-yl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 4-(1)-(2-isopropylthiazol-4-yl)methanone (Example 90, step g) (0.31 g). Days-Martin periodinane treatment (0.273 g) and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.025 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to (R)-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]pyrazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.169 g) and acetic acid (0.025 mL) in methanol (15 mL). The mixture was cooled in an ice-bath and treated with sodium cyanoborohydride (0.054 g). The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.125 g.

m/z 698(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D6-DMSO, 90 ° C) δ 11.25 (s, 1H), 7.93 (s, 1H), 7.56-7.46 (m, 2H), 7.31 - 7.25 (m, 1H), 6.89 (d) , J=8.5 Hz, 1H), 6.75 (d, J=8.5 Hz, 1H), 4.97-4.91 (m, 1H), 4.31 (s, 2H), 3.70 (s, 4H), 3.65 (s, 2H) , 3.50-3.37 (m, 2H), 3.33 - 3.26 (m, 1H), 3.23 - 3.15 (m, 2H), 3.12-3.02 (m, 4H), 2.06-1.97 (m, 2H), 1.83-1.72 ( m, 2H), 1.50 (s, 6H), 1.34 (d, J = 6.8 Hz, 6H). Five unobserved exchangeable protons.

Example 91

(R)-7-(2-(3-((4-(5-ethylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)methyl)-2-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (5-ethylthiophen-3-yl)(9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] Undecyl-4-yl)methanone

HATU (0.316g) was added to a 9- (3- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethyl) -2-fluoro-benzyl) -l-oxa-4,9- Diazaspiro[5.5]undecane (Example 78, step d) (0.271 g) and 5-ethylthiophene-3-carboxylic acid (0.1 g) and triethylamine (0.36 mL) in DMF (10 mL) Medium in the solution at 0 °C. The mixture was then stirred at 20 ° C for 1 hour. The reaction mixture was partitioned between EtOAc EtOAc. The residue was dissolved in EtOAc (20 mL)EtOAcEtOAc The reaction mixture was allowed to stand at 20 ° C for 18 hours and the solvent was removed under reduced pressure. The crude product was purified by flash chromatography on silica gel using 2% methanol in dichloromethane containing 1% triethylamine. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.12 g.

m/z 447(M+H) ⁺ (APCI)

b) (R)-7-(2-(3-((4-(5-ethylthiophen-3-carbonyl)-1-oxa-4,9-diazaza) Spiro[5.5]undecane-9-yl)methyl)-2-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzene well [d]thiazole-2(3H)-one Di-trifluoroacetate

(5-ethylthiophen-3-yl)(9-(2-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] eleven A solution of the carboxane-4-yl)methanone (Example 91, step a) (0.12 g) in DCM (15 mL) 0.171 g) and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.015 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.106 g) and acetic acid (0.015 mL) in methanol (15 mL). The mixture was cooled in an ice-bath and treated with sodium <RTI ID=0.0> The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.095 g.

m/z 655(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 11.27 (s, 1H), 7.52-7.41 (m, 3H), 7.25 (t, J = 7.7 Hz, 1H), 6.94 (d, J = 8.5 Hz, 1H), 6.89 (s, 1H), 6.77 (d, J = 8.2 Hz, 1H), 4.95-4.89 (m, 1H), 4.30 (s, 2H), 3.67 (t, J = 5.0 Hz, 2H), 3.53 (t, J = 5.0 Hz, 2H), 3.45 (s, 2H), 3.24 (t, J = 7.9 Hz, 2H), 3.18-3.03 (m, 8H), 2.81 (q, J = 7.5) Hz, 2H), 2.05-1.97 (m, 2H), 1.79-1.68 (m, 2H), 1.25 (t, J = 7.6 Hz, 3H). Five unobserved exchangeable protons.

Example 92

(R)-5-(2-(2-(2-isopropyl-3-(thiazolidine)-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenyl)-2-methylpropylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-three Fluoroacetate

a) (R)-5-(1-(Third-butyldimethylmethylalkyloxy)-2-(2-(2-fluoro-3-((4-(2-isopropylthiazole)- 4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenyl)-2-methylpropylamino)ethyl)- 8-hydroxyquinoline-2(1H)-one

(9-(2-Fluoro-3-(1-hydroxy-2-methylpropan-2-yl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 4-(1)-(2-Isopropylthiazol-4-yl)methanone (Example 90, Step g) (0.21 g) EtOAc. Days-Martin periodinane treatment (0.236 g) and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.025 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3mL) and added to (R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8-hydroxy-quinolin-2 (1H)-ketone (WO 2004106333) (0.186 g) in MeOH (15 mL). The mixture was cooled to 0 ° C and sodium triethoxysulfonate (0.136 g) was added in one portion. The reaction mixture was stirred at 20 ° C for 3 hours. The solvent was removed under reduced pressure and the residue was crystalljjjjjjjjjjjjjjj The crude product was purified by flash chromatography on EtOAc (EtOAc) eluting EtOAc EtOAc The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.28 g.

m/z 806(M+H) ⁺ (APCI)

b) (R)-5-(2-(2-(2-isopropyl-3-(thiazolidine-4-carbonyl)-1-oxa-4,9-diaza) Spiro[5.5]undecane-9-yl)methyl)phenyl)-2-methylpropylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one -trifluoroacetate

Triethylamine trihydrofluoride (0.074 mL) was added to methanol (2mL) to (R) -5- (1- (third - butyldimethylsilyl silicon alkyloxy) -2- (2- ( 2-fluoro-3-((4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl a solution of phenyl)-2-methylpropylamino)ethyl)-8-hydroxyquinolin-2(1H)-one (Example 92, step a) (0.28 g) in THF (8 mL) The reaction mixture was allowed to stand at 20 ° C for 18 hours. The solvent was removed under reduced pressure and the crude material was purified by preparative HPLC (Sunfire ^(s) , gradient: 10 to 40% acetonitrile in 0.2% aqueous TFA). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.175 g.

m/z 692(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.14 (d, J = 10.0 Hz, 1H), 7.93 (s, 1H), 7.57-7.50 (m, 2H), 7.29 (t, J = 7.8 Hz, 1H), 7.08 (d, J = 8.2 Hz, 1H), 6.98 (d, J = 8.2 Hz, 1H), 6.52 (d, J = 10.0 Hz, 1H), 5.40 - 5.34 (m, 1H) , 4.34 (s, 2H), 3.69 (s, 4H), 3.65 (s, 2H), 3.56-3.43 (m, 2H), 3.33-3.18 (m, 3H), 3.15-3.06 (m, 4H), 2.06 -1.98 (m, 2H), 1.83-1.72 (m, 2H), 1.52 (s, 6H), 1.34 (d, J = 7.8 Hz, 6H). Six unobserved exchangeable protons.

Example 93

(R)-7-(2-(3-((4-(5-ethylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)methyl)-5-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (5-ethylthiophen-3-yl)(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] Undecyl-4-yl)methanone

HATU (0.449 g) was added in one portion to 2-(3-(1-oxa-4,9-diazaspiro[5.5]undecal-9-ylmethyl)-5-fluorophenyl)ethanol ( Example 55, step c) (0.28 g) and 5-ethylthiophene-3-carboxylic acid (0.142 g) and triethylamine (0.506 mL) in EtOAc (EtOAc) The mixture was then stirred at 20 ° C for 1 hour. The reaction mixture was partitioned between EtOAc EtOAc. The crude product was purified by flash chromatography on silica gel using 2% methanol in dichloromethane containing 1% triethylamine. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.21 g.

m / z 447 (M + H ) + (APCI)

b) (R)-7-(2-(3-((4-(5-ethylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane) -9-yl)methyl)-5-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

(5-ethylthiophen-3-yl)(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] eleven A solution of the carboxane-4-yl)methanone (Example 93, step a) (0.21 g) in DCM (15 mL) g) Treatment, the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. Acetic acid (0.027 mL) was added to this solution, followed by removal of the solvent under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride A solution of the salt (WO 2007027134, Example 1, step d) (0.185 g) and acetic acid (0.027 mL) in methanol (15 mL). The mixture was cooled in an ice bath and treated with sodium cyanoborohydride (0.059 g). The cooling bath was removed, and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^(TM) , gradient: 10 to 40% acetonitrile in 0.2% aqueous TFA). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.115 g.

m/z 655(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.26 (s, 1H), 7.47 (s, 1H), 7.28-7.17 (m, 3H), 6.93 (d, J = 8.7Hz, 1H) , 6.89 (s, 1H), 6.77 (d, J = 8.2 Hz, 1H), 4.95-4.90 (m, 1H), 4.28 (s, 2H), 3.67 (t, J = 5.0 Hz, 2H), 3.53 ( t, J = 4.9 Hz, 2H), 3.45 (s, 2H), 3.27 (t, J = 7.9 Hz, 2H), 3.19 - 3.00 (m, 8H), 2.81 (q, J = 7.6 Hz, 2H), 2.07-1.99 (m, 2H), 1.80-1.68 (m, 2H), 1.25 (t, J = 7.4 Hz, 3H). Five unobserved exchangeable protons.

Example 94

( R )-8-Hydroxy-5-(1-hydroxy-2-(3-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]+monoalkan-9-yl)ethoxy)benzylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

3-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]+monoalkan-9-yl)ethoxy) benzaldehyde (example 10, step b) (0.24g) was added to (R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8- A mixture of hydroxyquinolin-2(1H)-one (WO 2004106333) (0.187 g) and acetic acid (0.032 mL) in methanol (2 mL). The mixture was stirred for 30 min and then cooled in an ice bath. Sodium triethoxysulfonium borohydride (0.178 g) was then added and the mixture was stirred for 2h then concentrated in vacuo. The residue was partitioned between ethyl acetate (50 mL) and EtOAc (EtOAc) The aqueous solution was separated and extracted with ethyl acetate (2×50 mL). The combined organic solution was washed with brine (20 mL) dried over sodium sulfate. Purification by gel chromatography, eluting with a gradient of 95:5:0.5 to 92:8:0.8 DCM:methanol: '880' aqueous ammonia solution. The fractions containing the product were combined, evaporated in vacuo and dissolved in THF (5 mL). Triethylamine trihydrofluoride (0.091 mL) was added and the mixture was stirred overnight. The solvent was evaporated in vacuo and the residue was dissolved in EtOAc EtOAc EtOAc Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile). The title compound was obtained as a white solid. The yield was 0.15 g.

m/z 634(M+H) ⁺ (APCI)

¹ H NMR (300 MHz, D ₆ -DMSO, 90 ° C) δ 8.09 (d, J = 10.0 Hz, 1H), 7.92 (s, 1H), 7.37 (t, J = 7.8 Hz, 1H), 7.20-6.94 (m, 5H), 6.50 (d, J = 10.0 Hz, 1H), 5.37 (dd, J = 8.2, 4.5 Hz, 1H), 4.37 (t, J = 5.0 Hz, 2H), 4.30-4.16 (m, 2H), 3.76-3.62 (m, 6H), 3.61-3.54 (m, 2H), 3.46-3.35 (m, 2H), 3.31-3.17 (m, 2H), 3.11-3.01 (m, 2H), 2.68 ( s, 3H), 2.11 - 1.98 (m, 2H), 1.94-1.78 (m, 2H). Six unobserved exchangeable protons.

Example 95

(R)-8-hydroxy-5-(1-hydroxy-2-(3-(3-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)propoxy)benzylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

a) 3-(3-Mexylphenoxy)propyl methanesulfonate

3-Bromo-1-propanol (3.94 mL) and potassium carbonate (6.22 g) were added to 3-hydroxybenzaldehyde (5 g) in acetonitrile (100 mL). The resulting mixture was stirred at reflux under nitrogen for 5 h. The reaction mixture was cooled to room temperature and partitioned between EtOAc and EtOAc. The organic layer was washed with brine, dried over sodium sulfate, filtered and evaporated. The residue was dissolved in DCM (30 mL The solution was cooled to 0 ° C and treated with methane sulfonium chloride (3.2 mL). The reaction mixture was stirred at 0 ° C for 10 minutes and then at room temperature for 1 hour. The mixture was washed twice with brine, dried over sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 5.20 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 9.98 (s, 1H), 7.50-7.43 (m, 2H), 7.40-7.39 (m, 1H), 7.20-7.16 (m, 1H), 4.46 (t, J = 6.2 Hz, 2H), 4.17 (t, J = 5.9 Hz, 2H), 3.01 (s, 3H), 2.30-2.23 (m, 2H).

b) 3-(3-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)propoxy Benzaldehyde

3-(3-Mexylphenoxy)propyl methanesulfonate (Example 95, Step a) (0.209 g) and (2-methylthiazol-4-yl)(1-oxa-4,9 -Diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 4, step h) (0.32 g) and triethylamine (0.282 mL) in acetonitrile (10 mL) The solution was heated at 65 ° C for 18 hours. The solvent was evaporated under reduced pressure and the residue was partitioned between ethyl acetate and brine. The organic layer was dried with sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.157 g.

m/z 444(M+H) ⁺ (APCI)

c) (R)-5-(1-( third -butyldimethylmethylalkyloxy)-2-(3-(3-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)propoxy)benzylamino)ethyl)-8-hydroxyquinoline-2(1H)-one

Sodium triethoxy borohydride (0.113 g) was added in one portion to 3-(3-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5 ] undec-9-yl) propoxy) benzaldehyde (example 95, step b) (0.157g), (R ) -5- (2- amino-l- (- butyldi Methyl decyloxy)ethyl)-8-hydroxyquinolin-2(1H)-one (WO 2004106333) (0.118 g) and acetic acid (0.020 mL) in methanol (7 mL) in a stirred solution at 0 ° C . The resulting mixture was stirred at 20 ° C for 2 hours. Most of the methanol was evaporated under reduced pressure and the residue was evaporated mjjjjjjjjjjjjj The crude product was purified by flash chromatography on EtOAc (EtOAc) eluting with EtOAc EtOAc The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.160 g.

m/z 762(M+H) ⁺ (APCI)

d) (R)-8-hydroxy-5-(1-hydroxy-2-(3-(3-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)propoxy)benzylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

( R )-5-(1-( Third -butyldimethylmethylalkyloxy)-2-(3-(3-(4-(2-methylthiazole-4-carbonyl)-1-oxyl) Hetero-4,9-diazaspiro[5.5]undecal-9-yl)propoxy)benzylamino)ethyl)-8-hydroxyquinoline-2(1H)-one (Example 95, Step c) (0.160 g) <RTI ID=0.0></RTI> The solvent was evaporated and the crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 5-35% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.081 mg.

m/z 648(M+H) ⁺ (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ10.75-10.43 (m, 2H), 10.00-9.74 (m, 1H), 9.30 (s, 1H), 9.10 (s, 1H), 8.07 (d, J = 9.7 Hz, 1H), 8.01 (s, 1H), 7.36 (t, J = 7.9 Hz, 1H), 7.16-7.09 (m, 3H), 7.01-6.94 (m, 2H), 6.54 (d, J = 9.7 Hz, 1H), 5.35 (d, J = 9.5 Hz, 1H), 4.21 (s, 2H), 4.07-4.00 (m, 2H), 3.83-3.16 (m, 10H), 3.08-2.90 (m, 4H) ), 2.70 (s, 3H), 2.18-2.04 (m, 4H), 1.83-1.66 (m, 2H). An unobserved exchangeable proton.

Example 96

(R)-8-hydroxy-5-(1-hydroxy-2-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)benzylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

a) 3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl)benzaldehyde

(2-methylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 4, step h (0.408 g), a solution of 3-(bromomethyl)benzaldehyde (0.205 g) and triethylamine (0.36 mL) in EtOAc (10 mL) The solution was concentrated in vacuo. The organic phase was washed twice with water, EtOAc (EtOAc) EtOAc. Things. The yield was 0.325 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 10.01 (s, 1H), 7.85 (d, J = 0.8 Hz, 1H), 7.80 (s, 1H), 7.76 (d, J = 7.4 Hz , 1H), 7.61 (d, J = 7.4 Hz, 1H), 7.52 (t, J = 7.4 Hz, 1H), 3.71-3.46 (m, 8H), 2.68 (s, 3H), 2.44 - 2.29 (m, 4H), 1.78-1.65 (m, 2H), 1.61-1.46 (m, 2H).

b) (R)-5-(1-( third -butyldimethylmethylalkyloxy)-2-(3-((4-(2-) Thiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)benzylamino)ethyl)-8-hydroxyquinoline -2(1H)-ketone

3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl)benzaldehyde (implementation) Example 96, step a) (0.303g), (R ) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8-hydroxy-quinolin-2 A solution of (1H)-ketone (WO 2004106333) (0.330 g) and acetic acid (0.043 mL) in methanol (5 mL) was stirred at room temperature for 30 min then cooled in ice-water bath under nitrogen and triethylamine Treatment with sodium oxyborohydride (0.243 g). The mixture was stirred in ice-water for 2 h then concentrated in vacuo. The residue was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate and partitioned, and the aqueous phase was extracted twice with ethyl acetate. Any insoluble gum residue is dissolved in methanol. The combined ethyl acetate phase was washed with brine, dried over magnesium sulfate and combined with methanol. Purification by flash chromatography on ruthenium chloride eluting with 1:7:92 triethylamine:methanol:dichloromethane afforded a yellow foam of slightly sub-subtitle product. The yield was 0.324 g.

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.35 (d, J = 10.0 Hz, 1H), 8.1 (s, 1H), 7.40-7.30 (m, 2H), 7.30-7.23 (m, 2H), 7.17 (d, J = 8.2 Hz, 1H), 7.08 (d, J = 8.2 Hz, 1H), 6.59 (d, J = 10.0 Hz, 1H), 5.30 (dd, J = 7.3, 4.7 Hz, 1H), 3.91-3.70 (m, 8H), 3.57 (s, 2H), 3.02 (dd, J = 12.3, 7.4 Hz, 1H), 2.91-2.77 (m, 4H), 2.56-2.43 (m, 4H) , 1.90-1.80 (m, 2H), 1.74-1.62 (m, 2H), 0.99 (s, 9H), 0.19 (s, 3H), 0.00 (s, 3H). Three unobserved exchangeable protons.

c) (R)-8-hydroxy-5-(1-hydroxy-2-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaza) Spiro[5.5]undecane-9-yl)methyl)benzylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

( R )-5-(1-(Third-butyldimethylmethylalkyloxy)-2-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-) 4,9-diazaspiro[5.5]undecane-9-yl)methyl)benzylamino)ethyl)-8-hydroxyquinolin-2(1H)-one (Example 96, step b) (0.320 g) and a solution of triethylamine trihydrofluoride (0.15 mL) in THF (5 mL). The residue was dissolved in acetonitrile (3ml) and water (1ml) of the mixture, and the formed purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile) was borrowed. The product containing fractions were concentrated in vacuo and co-evaporated three times from acetonitrile to yield a white foam. The foam was triturated with diethyl ether to give a solid, which was crystallised, washed with diethyl ether and dried at room temperature in vacuo to give the title compound as white powder. The yield was 0.264 g.

m/z 604(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.08 (d, J = 9.7 Hz, 1H), 7.90 (s, 1H), 7.65-7.49 (m, 4H), 7.10 (d, J = 7.9 Hz, 1H), 6.97 (d, J = 8.2 Hz, 1H), 6.52 (d, J = 10.0 Hz, 1H), 5.35 (dd, J = 8.6, 4.2 Hz, 1H), 4.35 - 4.21 (m, 4H), 3.74-3.57 (m, 6H), 3.23-2.98 (m, 6H), 2.67 (s, 3H), 2.06-1.91 (m, 2H), 1.85-1.65 (m, 2H). Six unobserved exchangeable protons.

Example 97

(R)-5-(2-(2,5-Dimethyl-4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)ethoxy)benzylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetate

2,5-Dimethyl-4-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 9- yl) ethoxy) benzaldehyde (example 18, step f) (0.08g) was added to (R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy A mixture of ethyl)ethyl)-8-hydroxyquinolin-2(1H)-one (WO 2004106333) (0.088 g) and acetic acid (0.010 mL) in methanol (2 mL). The mixture was stirred for 30 min and then cooled in an ice bath. Sodium cyanoborohydride (0.016 g) was then added and the mixture was stirred for 2 h. The solvent was removed in vacuo and the residue was purified eluting with EtOAc EtOAc (EtOAc:EtOAc:EtOAc The fractions containing the product were combined and evaporated in vacuo. The residue was redissolved in tetrahydrofuran (5 mL), triethylamine tribr. The solvent was evaporated in vacuo. Purification by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 5 to 50% acetonitrile). The title compound was obtained as a white solid. The yield was 0.07 g.

m/z 662(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.12 (d, J = 10.0 Hz, 1H), 7.92 (s, 1H), 7.27 (s, 1H), 7.12 (d, J = 8.2 Hz) , 1H), 6.99 (d, J = 8.2 Hz, 1H), 6.86 (s, 1H), 6.51 (d, J = 9.7 Hz, 1H), 5.38 (dd, J = 8.2, 4.6 Hz, 1H), 4.35 (t, J = 4.9 Hz, 2H), 4.25-4.13 (m, 2H), 3.76-3.64 (m, 6H), 3.61-3.53 (m, 2H), 3.46-3.36 (m, 2H), 3.33-3.20 (m, 2H), 3.19-3.10 (m, 2H), 2.68 (s, 3H), 2.33 (s, 3H), 2.15 (s, 3H), 2.12-2.01 (m, 2H), 1.95-1.79 (m , 2H). Six unobserved exchangeable protons.

Example 98

(R)-8-hydroxy-5-(1-hydroxy-2-((5-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diaza) Heterospiro[5.5]undecane-9-yl)ethyl)thiophen-2-yl)methylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

a)(R)-5-(1-( third -butyldimethylmethylalkyloxy)-2-((5-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9- Ethyl)thiophen-2-yl)methylamino)ethyl)-8-hydroxyquinolin-2(1H)-one di-trifluoroacetate

5-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethyl)thiophene- 2- carbaldehyde (example 21, step c) (0.188g) and (R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8- A solution of hydroxyquinolin-2(1H)-one (WO 2004106333) (0.150 g) in MeOH (10 mL) was taken fromEtOAc (EtOAc) The mixture was stirred at 20 ° C for 18 hours. Additional sodium triethoxysulfonate (0.143 g) was added and stirring was continued at 20 °C for 2 hours. Additional sodium triethoxysulfonate (143 mg) was added and stirred for 2 hours. Methanol was removed under reduced pressure and the residue was partitioned between EtOAc EtOAc. The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.122 g.

m/z 738(M+H) ⁺ (APCI)

b) (R)-8-hydroxy-5-(1-hydroxy-2-((5-(2-(4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-) Diazaspiro[5.5]undecane-9-yl)ethyl)thiophen-2-yl)methylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

( R )-5-(1-( Third -butyldimethylmethylalkyloxy)-2-((5-(2-(4-(2-methylthiazole-4-carbonyl)-1-) Oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethyl)thiophen-2-yl)methylamino)ethyl)-8-hydroxyquinoline-2 (1H a solution of the ketone (Example 98, step a) (0.122 g) in EtOAc (EtOAc) (EtOAc) The mixture was placed at 20 ° C for 18 hours. The solvent was removed under reduced pressure and the crude material was purified by preparative HPLC (Sunfire ^(s) , gradient: 10 to 40% acetonitrile in 0.2% aqueous TFA). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.085 g.

m/z 624(M+H) ⁺ (APCI)

¹ H NMR (400 MHz, D ₆ -DMSO, 90 ° C) δ 8.10 (d, J = 31.8 Hz, 1H), 7.93 (s, 1H), 7.15 (d, J = 12.5 Hz, 1H), 7.10 (d) , J = 15.7 Hz, 1H), 6.98 (d, J = 22.6 Hz, 1H), 6.92 (d, J = 11.2 Hz, 1H), 6.53 (d, J = 30.4 Hz, 1H), 5.37 - 5.31 (m , 1H), 4.41 (dd, J = 17.7, 14.4 Hz, 2H), 3.71 (s, 4H), 3.66 (s, 2H), 3.42-3.33 (m, 4H), 3.27-3.06 (m, 6H), 2.68 (s, 3H), 2.10 - 1.98 (m, 2H), 1.88-1.75 (m, 2H). Six unobserved exchangeable protons.

Example 99

(R)-7-(2-(5-((4-(5-ethylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)methyl)-2-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (5-ethylthiophen-3-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate

HATU (1.15g) was added in one portion to 1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester hydrochloride (WuXi PharmaTech) (0.682 g) and 5- Ethylthiophene-3-carboxylic acid (0.364 g) and triethylamine (1.3 mL) were dissolved in EtOAc (EtOAc)EtOAc. The mixture was then stirred at 20 ° C for 1 hour. The reaction mixture was partitioned between EtOAc EtOAc. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure soluble fractions were evaporated to dryness to give a &lt;RTI ID=0.0&gt; DCM (10 mL) was added followed by trifluoroacetic acid (10 mL) and the resulting solution was allowed to stand at 20 ° C for 25 min. Toluene (30 mL) was added and the solvent was evaporated under reduced pressure. The residue was azeotroped twice with acetonitrile to give the subtitle compound. The yield was 0.950 g.

m/z 295(M+H) ⁺ (APCI)

b) (5-ethylthiophen-3-yl)(9-(4-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxyl Hetero-4,9-diazaspiro[5.5]undecane-4-yl)methanone

2-(5-(Bromomethyl)-2-fluorophenyl)ethanol (Example 47, step a) (0.171 g) was added to (5-ethylthiophen-3-yl)(1-oxo- 4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 99, step a) (0.3 g) and triethylamine (0.410 ml) in acetonitrile (15 mL) The solution was stirred and the mixture was stirred at 20 ° C for 2 hours. The solvent was evaporated under reduced pressure. The organic layer was dried with sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on silica gel using 1% triethylamine and 2% methanol in dichloromethane. The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.2 g.

m/z 447(M+H) ⁺ (APCI)

c) (R)-7-(2-(5-((4-(5-ethylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane -9-yl)methyl)-2-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

(5-ethylthiophen-3-yl)(9-(4-fluoro-3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] eleven A solution of the carboxane-4-yl)methanone (Example 99, step b) (0.2 g) in DCM (15 mL) 0.285 g) and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.026 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.176 g) and acetic acid (0.026 mL) in methanol (15 mL). The mixture was cooled in an ice-bath and treated with sodium cyanoborohydride (0.056 g). The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.14 g.

(M+H) ⁺ 655 (APCI)

_{^{1 H NMR (400MHz, D 6}} -DMSO, 90 ℃) δ11.27 (s, 1H), 7.49-7.44 (m, 3H), 7.29-7.23 (m, 1H), 6.94 (d, J = 7.2Hz, 1H), 6.89 (s, 1H), 6.77 (d, J = 8.2 Hz, 1H), 4.94-4.89 (m, 1H), 4.26 (s, 2H), 3.67 (t, J = 5.0 Hz, 2H), 3.53 (t, J = 5.0 Hz, 2H), 3.45 (s, 2H), 3.24 (t, J = 8.1 Hz, 2H), 3.18-3.02 (m, 8H), 2.81 (q, J = 7.5 Hz, 2H) ), 2.07-1.98 (m, 2H), 1.79-1.67 (m, 2H), 1.25 (t, J = 7.4 Hz, 3H). Five unobserved exchangeable protons.

Example 100

( R - 7-(2-(3-((4-(5-ethylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl) )methyl)-4-fluorophenethylamino)-1-indolylethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) tert-butyl(4-fluorophenylethoxy)dimethyl decane

Third -butyl dimethyl decyl chloride (0.903 g) was added in portions to 2-(4-fluorophenyl)ethanol (0.7 g) and imidazole (0.408 g) in DMF (20 mL) stirred at 20 ° C In solution. The reaction mixture was stirred at room temperature for 3 hours and then partitioned between ethyl acetate and brine. The organic layer was washed twice with brine, dried over sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.99 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.18-7.13 (m, 2H), 6.99-6.93 (m, 2H), 3.77 (t, J = 6.9Hz, 2H), 2.78 (t, J = 6.8Hz, 2H), 0.86 (s, 9H), -0.03 (s, 6H)

b) 5-(2-( third -butyldimethylmethylalkyloxy)ethyl)-2-fluorobenzaldehyde

The second butyl lithium (1.4 M in cyclohexane, 2.78 ml) was added to dry THF (10 mL) under nitrogen and the solution was cooled to -78. N1-(2-(Dimethylamino)ethyl)-N1 ,N2,N2 -trimethylethane-1,2-diamine (0.674 g) was slowly added dropwise. Then added dropwise over 5 minutes to a third - butyl (4-fluorophenyl ethoxy) Dimethyl Silane (Example 100, step a) (0.99g) in dry tetrahydrofuran, the (3mL) was added. The reaction mixture was stirred at -78 °C for 2 hours. DMF (2.1 mL) was added dropwise over 5 min and the mixture was stirred at -78 °C for one hour and then at room temperature for 45 min. The reaction mixture was quenched by the addition of water. The acid ethyl ester (200 mL) was added and the organics were washed three times with water then twice with 2M HCl and then twice with water. The organic solution was washed with brine, dried over sodium sulfate, filtered and evaporated. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield is 0.5g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 10.36 (s, 1H), 7.72-7.69 (m, 1H), 7.48-7.43 (m, 1H), 7.11-7.06 (m, 1H), 3.80 (t, J = 6.5 Hz, 2H), 2.83 (t, J = 6.4 Hz, 2H), 0.85 (s, 9H), 0.04 (s, 6H).

c) (5-ethylthiophen-3-yl)(9-(2-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa- 4 ,9-diazaspiro[5.5]undecane-4-yl)methanone

Add sodium triethoxysulfonate (0.225 g) to (5-ethylthiophen-3-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4- ethyl) -2-fluorobenzaldehyde (- yl) methanone trifluoroacetate (butyldimethyl silicon alkyloxy step a) (0.289g) and 5- (2- (third embodiment 99) Example 100, step b) (0.2 g) and acetic acid (0.041 ml) in NMP (20 mL). The reaction mixture was then stirred at 20 ° C for 18 hours. Additional sodium triethoxysulfonate (0.150 g) was added and stirred for 4 hours. The mixture was partitioned between EtOAc and EtOAc (EtOAc)EtOAc. The resulting gum was dissolved in THF (20 mL) and taken to EtOAc (1M, EtOAc. The solution was allowed to stand at 20 ° C for 8 hours. The solvent was evaporated under reduced pressure and the crude material was purified eluted eluted elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.18 g.

m/z 447(M+H) ⁺ (APCI)

d) (R)-7-(2-(3-((4-(5-ethylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane -9-yl)methyl)-4-fluorophenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

(5-ethylthiophen-3-yl)(9-(2-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] eleven A solution of the carboxane-4-yl)methanone (Example 100, step c) (0.18 g) in DCM (15 mL) 0.256 g) and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. sodium sulphate (20 mL) and EtOAc (EtOAc) The mixture was extracted twice with ethyl acetate. EtOAc (EtOAc)EtOAc. Acetic acid (0.023 mL) was added to this solution and the solvent was removed under reduced pressure. The residue was dissolved in methanol (3 mL) and added to ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.159 g) and acetic acid (0.023 mL) in methanol (15 mL). The mixture was cooled in an ice-bath and treated with sodium cyanoborohydride (0.051 g). The cooling bath was removed and the mixture was stirred at 20 ° C for 3 hours. The solvent was evaporated under reduced pressure to a volume of 3 mL and THF (20 mL). The mixture was washed with a mixture of saturated brine (10 mL) and saturated sodium hydrogen sulfate (1 mL). The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The residue was azeotroped twice with acetonitrile. The crude product was purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in 10 to 40% acetonitrile). The fractions containing the desired compound are evaporated to dryness to give the title compound. The yield was 0.14 g.

m/z 655(M+H) ⁺ (APCI)

^{1 H NMR (400MHz, D6-} DMSO, 90 ℃) δ11.27 (s, 1H), 7.48-7.43 (m, 2H), 7.42-7.37 (m, 1H), 7.25 (t, J = 9.1Hz, 1H ), 6.93 (d, J = 8.1 Hz, 1H), 6.89 (s, 1H), 6.77 (d, J = 8.5 Hz, 1H), 4.94-4.88 (m, 1H), 4.27 (s, 2H), 3.67 (t, J = 5.0 Hz, 2H), 3.53 (t, J = 5.0 Hz, 2H), 3.45 (s, 2H), 3.23 (t, J = 8.2 Hz, 2H), 3.18-2.97 (m, 8H) , 2.81 (q, J = 7.8 Hz, 2H), 2.04-1.98 (m, 2H), 1.79-1.69 (m, 2H), 1.25 (t, J = 7.6 Hz, 3H). Five unobserved exchangeable protons.

Example 101

(R)-4-hydroxy-7-(1-hydroxy-2-(2-(6-(4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diaza) Heterospiro[5.5]undecane-9-yl)hexylthio)ethylamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

a) 6-(2,2-dimethoxyethylthio)hexane-1-ol

A solution of 6-mercapto-hexane-l-ol (2.5 mL) in MeCN (30 mL) EtOAc (EtOAc:EtOAc The mixture was stirred at 0 ° C for 1 h then 2-bromo-1,1-dimethoxy-ethane (2.4 mL). The resulting mixture was stirred at RT for 20 h then quenched by addition of saturated sodium bicarbonate. The mixture was extracted with EtOAc (EtOAc) (EtOAc)EtOAc. Purification by silica gel chromatography, eluting with 25% ethyl acetate in hexanes to give the subtitle compound as a colourless liquid. The yield was 1.85 g.

¹ H NMR (400 MHz, D _{4 -} MeOH) δ 4.49 (t, J = 5.5 Hz, 1H), 3.64 (t, J = 6.6 Hz, 2H), 3.37 (s, 6H), 2.69 (d, J = 5.5 Hz, 2H), 2.59 (t, J = 7.4 Hz, 2H), 1.66-1.52 (m, 4H), 1.46-1.32 (m, 4H). An unobserved exchangeable proton.

b) 2-(6-bromohexylthio)acetaldehyde

In the 6- (2,2-dimethoxy ethylthio) hexan-1-ol (Example 101, step a) (1.85g) in DCM (75mL) was added under N ₂ at 0 ℃ Carbon tetrabromide (3.31 g) was then added in portions to triphenylphosphine (2.62 g). The resulting mixture was stirred at RT for 1.25 h then concentrated in vacuo. Purification by gel chromatography eluting with 0 to 50% ethyl acetate in cyclohexane afforded the subtitle compound as a yellow liquid. The yield was 1.31 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 9.47 (t, J = 3.5 Hz, 1H), 3.43 - 3.36 (m, 2H), 3.18 (d, J = 3.5 Hz, 2H), 2.44 (t, J = 7.3 Hz, 2H), 1.91-1.81 (m, 2H), 1.64-1.53 (m, 2H), 1.49-1.36 (m, 4H).

c) (R)-2-(6-bromohexylthio)ethyl (2-hydroxy-2-(4-hydroxy-2-yloxy-2,3-dihydrobenzo[d]thiazole-7 -ethyl)ethyl)amino carboxylic acid tert-butyl ester

a solution of 2-(6-bromohexylthio)acetaldehyde (Example 101, step b) (1.2 g) in DMF (20 mL) Molecular sieves ( R )-7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride was added at 0 ° C under N ₂ (WO 2007027134 , Example 1, step d) (1.45 g). The resulting mixture was stirred at 0&lt;0&gt;C for 1 h then EtOAc (EtOAc &lt Di-tert-butyl dicarbonate (1.1 g) was added and stirring was continued for 20 h. The reaction mixture was quenched by the addition of a saturated sodium hydrogen carbonate solution (40 mL). The combined organic solution was washed with brine, dried over sodium sulfate, filtered and evaporated in vacuo. Purification by chromatography on silica gel eluting with EtOAc EtOAc (EtOAc) The yield was 0.56 g.

¹ H NMR (300 MHz, D _{4 -} MeOH) δ 6.89 (dd, J = 18.6, 8.3 Hz, 1H), 6.72 (d, J = 8.2 Hz, 1H), 4.87 (s, 1H), 3.59-3.39 (m) , 4H), 3.33-3.29 (m, 2H), 2.64-2.41 (m, 4H), 1.89-1.80 (m, 2H), 1.66-1.32 (m, 15H). Three unobserved exchangeable protons.

d) 4-(2,2,2-trifluoroethenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester

A solution of trifluoroacetic anhydride (2.2 mL) in DCM (14 mL) was added dropwise over a period of 20 min to 1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert. The ester hydrochloride (WuXi PharmaTech) (4.7 g) and triethylamine (4.9 mL) were taken in DCM (90 mL) in EtOAc. The mixture was stirred at 0 ° C for 45 minutes and then with RT for 2 hours. Water (90 mL) was added and the mixture was stirred vigorously for 10 min. The layers were separated and the aqueous layer was extracted with DCM (50mL). The combined organic layers were dried over sodium sulfate and evaporated in vacuo. The residue was purified by EtOAc EtOAc (EtOAc) The yield was 2.73 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ3.86 (d, J = 13.3Hz, 1H), 3.77 (t, J = 4.9Hz, 2H), 3.74-3.64 (m, 2H), 3.61 (t, J = 4.8 Hz, 1H), 3.52 (s, 1H), 3.23 - 3.15 (m, 1H), 3.07 (t, J = 11.4 Hz, 1H), 1.85 (d, J = 13.8 Hz, 1H), 1.77 (d, J = 13.9 Hz, 1H), 1.52-1.37 (m, 12H).

e) 2,2,2-trifluoro-1-(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)ethanone trifluoroacetate

Adding trifluoroacetic acid (66 mL) to 4-(2,2,2-trifluoroethenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid Tributyl ester (Example 101, step d) (4.33 g) in EtOAc m. Toluene (50 mL) was added and the mixture was evaporated (EtOAc) elut The yield was 5.52 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ8.73 (brs, 2H), 3.77 (d, J = 5.1Hz, 2H), 3.72 (d, J = 5.3Hz, 1H), 3.65 (d, J = 5.0Hz , 1H), 3.58 (s, 1H), 3.48 (s, 1H), 3.33 (s, 2H), 3.24 (s, 2H), 2.08 (t, J = 14.6 Hz, 2H), 1.93-1.75 (m, 2H).

f) (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl (2-(6-(4) -(2,2,2-trifluoroethylidene)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)hexylthio)ethyl)aminocarboxylic acid Third butyl ester

(R)-2-(6-bromohexylthio)ethyl (2-hydroxy-2-(4-hydroxy-2-yloxy-2,3-dihydrobenzo[d]thiazole-7-yl) Ethyl) butyl methacrylate (Example 101, step c) (0.55 g) and 2,2,2-trifluoro-1-(1-oxa-4,9-diaza-snail [5.5] undec-4-yl)-ethanone trifluoroacetate (Example 101, step e) (0.25 g) and triethylamine (0.26 mL) combined in acetonitrile (20 mL) and heated at 80 ° C for 48 h . The volatiles were evaporated in vacuo. Purification was achieved by silica gel chromatography eluting with 10% MeOH in DCM Yield 0.10 g and 0.074 g (less pure).

m/z 721(M+H) ⁺

¹ H NMR (400 MHz, D _{4 -} MeOH) δ 6.81 (s, 1H), 6.69 (d, J = 8.21 Hz, 1H), 3.80-3.74 (m, 2H), 3.65 (s, 2H), 3.58- 3.54 (m, 1H), 3.52-3.47 (m, 1H), 3.11-2.94 (m, 4H), 2.51-2.41 (m, 6H), 2.05-1.95 (m, 2H), 1.80-1.59 (m, 8H) ), 1.50 (s, 9H), 1.46-1.25 (m, 6H). A proton that is obscured by the solvent peak and three unobserved exchangeable protons.

g) (R)-2-(6-(1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)hexylthio)ethyl (2-hydroxy-2- (4-Hydroxy-2-oxo-2,3-dihydrobenzene well [d]thiazole-7-yl)ethyl)carbamic acid tert-butyl ester

(R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl (2-(6-(4-() 2,2,2-Trifluoroethenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)hexylthio)ethyl)aminocarboxylic acid A solution of potassium carbonate (0.06 g) in water (10 mL) was added to a stirred solution of EtOAc (EtOAc). The mixture was stirred at RT for 17 h before removing the methanol under a stream of nitrogen. The solution was diluted with EtOAc (EtOAc) (EtOAc)EtOAc. The residue was dissolved in MeOH (EtOAc m. The yield was 0.15 g.

m/z 625(M+H) ⁺

¹ H NMR (300 MHz, D ₄ -MeOH) δ 6.94 - 6.82 (m, 1H), 6.73 (d, J = 2.0 Hz, 1H), 3.66 - 3.59 (m, 2H), 3.51-3.40 (m, 2H) ), 2.80-2.69 (m, 4H), 2, 66-2.62 (m, 2H), 2.55-2.40 (m, 8H), 2.04-1.94 (m, 2H), 1.62-1.56 (m, 8H), 1.55 -1.21 (m, 13H).

A proton that is obscured by the solvent peak and four unobserved exchangeable protons.

h) (R)-4-hydroxy-7-(1-hydroxy-2-(2-(6-(4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-) Diazaspiro[5.5]undecane-9-yl)hexylthio)ethylamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

(R)-2-(6-(1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)hexylthio)ethyl (2-hydroxy-2-(4) -Hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazole-7-yl)ethyl)carbamic acid tert-butyl ester (Example 101, step g) (0.093 g) in DMF Triethylamine (0.062 mL) and 2-isopropyl-thiazole-4-carboxylic acid (0.025 g) were added to the solution in (3 mL), followed by O-(7-azabenzotriazole-1- Base) -N,N,N',N'-tetramethyluronium (0.079 g). The resulting mixture was stirred at RT for 4 h, then 4 drops of &apos;880&quot; aqueous ammonia solution was added and the mixture was stirred for 15 min. Brine and ethyl acetate were added to the solution and the phases were separated, after which the organic phase was extracted with ethyl acetate (×2). The combined organic phases were dried over sodium sulfate, filtered and evaporated in vacuo. The residue was dissolved in DCM (2 mL). The solution was left at RT for 40 min then toluene (20 mL) was added and the mixture was concentrated in vacuo. An additional amount of toluene was added and the mixture was concentrated again, after which the material was azeotroped with acetonitrile. Purified by preparative HPLC (Phenomenex , Gradient: 10 to 40% acetonitrile in 0.1% aqueous formic acid). The product-containing fractions were combined and lyophilized to give the title compound as a colourless solid. The yield was 0.016 g.

m/z 678(M+H) ⁺

¹ H NMR (400 MHz, D ₆ -DMSO, 80 ° C) δ 8.18 (s, 1H), 7.91 (d, J = 0.7 Hz, 1H), 6.90 (d, J = 8.3 Hz, 1H), 6.72 (d) , J = 8.3 Hz, 1H), 4.64 - 4.58 (m, 1H), 3.72-3.62 (m, 2H), 3.62-3.58 (m, 6H), 3.38-3.28 (m, 1H), 2.84 - 2.74 (m ,6H),2.62-2.58(m,2H), 2.47-2.31(m,6H),1.78-1.69(m,2H),1.60-1.50(m,4H),1.45-1.41(m,2H),1.38 (d, J = 6.6 Hz, 6H), 1.32-1.26 (m, 2H). Five unobserved exchangeable protons.

Example 102

(R)-4-hydroxy-7-(1-hydroxy-2-(7-(4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)-2,2-dimethylheptylamino)ethyl)benzo[d]thiazole-2(3H)-one formate

a) 7-Bromo-2,2-dimethylheptanoic acid ethyl ester

A solution of diisopropylamine (5.82 mL) in THF (30 mL) was added n- butyllithium (2.5M, 16.4 mL in hexanes) and the mixture was stirred at this temperature for 30 min then cooled to -78 °C. Ethyl isobutyrate (5 mL) was added dropwise, and the resulting mixture was stirred at -78 &lt;0&gt;C for 1 h then EtOAc &lt The reaction mixture was stirred at -78 °C for 1 h, then at RT for 2.5 h then poured into saturated aqueous ammonium chloride. The solution was extracted with EtOAc (×2) then EtOAc (EtOAc) Purification by gel chromatography eluting with 0 to 25% ethyl acetate in cyclohexane afforded a sub-title compound as a yellow liquid. The yield was 4.45 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 4.08 (q, J = 7.1 Hz, 2H), 3.41-3.33 (m, 2H), 1.91-1.77 (m, 2H), 1.61-1.44 (m, 2H), 1.52-1.24 (m, 2H), 1.21 (t, J = 7.1 Hz, 5H), 1.12 (s, 6H).

b) 7-bromo-2,2-dimethylheptan-1-ol

7-bromo-2,2-dimethyl-heptanoic acid ethyl ester (Example 102, step a) (1.5g) in dry diethyl ether (50mL) at 0 ℃ in the solution was added dropwise under N ₂ for hydrogenation diisopropyl Butyl aluminum (1 M in toluene, 12.5 mL). The reaction mixture was stirred at 0&lt;0&gt;C for 1 h then quenched (150 mL). The mixture was stirred for 1 h then extracted with EtOAc (×3). The combined organics were washed with brine and dried over sodium sulfate, filtered and evaporated in vacuo. Purification by gel chromatography eluting with 0 to 25% ethyl acetate in cyclohexane afforded the subtitle compound as a colourless liquid. The yield was 1.16 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 3.41 (t, J = 6.8 Hz, 2H), 3.32 (d, J = 5.8 Hz, 2H), 1.93-1.82 (m, 2H), 1.47-1.37 (m, 2H), 1.35-1.20 (m, 4H), 0.91-0.81 (m, 6H). An unobserved exchangeable proton.

c) (9-(7-fluorenyl-6,6-dimethylheptyl)-1-oxa-4,9-diazaspiro[5.5] Undecyl-4-yl)(2-isopropylthiazol-4-yl)methanone

(2-Isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (0.5 g) was dissolved in methanol and applied to an SCX column pre-wetted with methanol. The column was washed with methanol and dissolved in 2M ammonia in methanol. The eluent evaporates in vacuo to yield the material as a free base (0.29 g). Add 7-bromo-2,2-dimethylheptan-1-ol (Example 102, step b) (0.26 g) in acetonitrile (10 mL) then triethylamine (0.27 mL). The resulting mixture was heated at 60 ° C for 17 h. The volatiles were evaporated in vacuo and EtOAc (EtOAc) elute. The yield was 0.29 g.

m/z 452(M+H) ⁺

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.87 (s, 1H), 4.01-3.95 (m, 2H), 3.80-3.62 (m, 2H), 3.49 (s, 1H), 3.34-3.28 (m, 2H) ), 3.16-3.09 (m, 1H), 2.98-2.75 (m, 3H), 2.42-2.20 (m, 2H), 2.09 (brm, 2H), 1.95-1.82 (m, 2H), 1.47-1.40 (m , 8H), 1.34-1.21 (m, 8H), 0.85 (s, 6H). An unobserved exchangeable proton.

d) 7-(4-(2-Isopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)-2,2- Dimethylheptanal

Trifluoroacetic acid (0.048 mL) was added to (9-(7-hydroxy-6,6-dimethylheptyl)-1-oxa-4,9-diazaspiro[5.5] under argon at 0 °C. a solution of the undecane-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 102, step c) (0.28 g) Dess-Martin periodinane (0.39 g) was then added. The reaction mixture was stirred at rt for 1 h then EtOAc EtOAc EtOAc EtOAc. The layers were separated and the aqueous solution was extracted with ethyl acetate (×2). The combined organic solution was washed with a saturated sodium bicarbonate solution. Acetic acid (0.053 mL) was added to EtOAc (EtOAc)EtOAc. The yield was 0.33 g.

m/z 450(M+H) ⁺

_{^{1 H NMR (300MHz, CDCl 3}} ) δ 9.43 (s, 1H), 7.86 (s, 1H), 4.02-3.86 (m, 3H), 3.82-3.67 (m, 4H), 3.36-3.26 (m, 2H) , 3.24 - 2.97 (m, 2H), 2.85-2.66 (m, 4H), 2.03 (d, J = 8.4 Hz, 2H), 1.79-1.60 (s, 2H), 1.48-1.35 (m, 8H), 1.34 -1.19 (m, 4H), 1.06 (s, 6H).

e) (R)-4-hydroxy-7-(1-hydroxy-2-(7-(4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)-2,2-dimethylheptylamino)ethyl)benzo[d]thiazole-2(3H)-one formate

Acetic acid (0.053 mL) was added to 7-(4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl -2,2-dimethylheptanal (Example 102, step d) (0.33 g) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d] Thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.25 g) and 3 The molecular sieves were in a mixture of anhydrous methanol (10 mL). The mixture was stirred for 5 min and then cooled to 0 °C. Sodium triethoxysulfonium borohydride (0.13 g) was added and the resulting mixture was stirred at RT for 16.5 h. The solution was filtered and concentrated in vacuo. The residue is dissolved in a mixture of acetonitrile and water and purified by preparative HPLC (Phenomenex , Gradient: 10 to 40% acetonitrile in 0.1% aqueous formic acid). The product-containing fractions were combined and lyophilized to give the title compound as a white solid. The yield is 0.1 g.

m/z 660(M+H) ⁺

¹ H NMR (400 MHz, D _{4 -} MeOH) δ 8.50 (s, 1H), 7.89 (s, 1H), 6.93 (d, J = 8.3 Hz, 1H), 6.71 (d, J = 8.3 Hz, 1H) , 4.93 (dd, J=9.5, 4.1 Hz, 1H), 3.92-3.56 (m, 6H), 3.37-3.26 (m, 1H), 3.10-2.66 (m, 10H), 2.11-1.96 (m, 2H) , 1.84-1.57 (m, 4H), 1.38 (d, J = 6.9 Hz, 6H), 1.29 (s, 6H), 0.96 (s, 6H). Five unobserved exchangeable protons.

Example 103

(R)-4-hydroxy-7-(1-hydroxy-2-(9-(4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)-2,2-dimethylhydrazinoamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

a) ethyl 9-bromo-2,2-dimethyl decanoate

A solution of diisopropylamine (5.82 mL) in THF (30 mL) was added n-butyllithium (2.5M, 16.4 mL in hexanes) and the mixture was stirred at this temperature for 30 min then cooled to -78 °C. Ethyl isobutyrate (5 mL) was added dropwise, and the resulting mixture was stirred at -78 &lt;0&gt;C for 1 h then EtOAc (EtOAc) The reaction mixture was stirred at -78 ° C for 1 h, then with RT for 2 h then poured into saturated aqueous ammonium chloride. The solution was extracted with EtOAc (EtOAc). Purification by gel chromatography eluting with 0 to 10% ethyl acetate in cyclohexane afforded the subtitle compound as a colourless liquid. The yield was 1.27 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ4.11 (q, J = 7.1Hz, 2H), 3.43-3.37 (m, 2H), 1.91-1.80 (m, 2H), 1.56-1.36 (m, 4H), 1.39-1.21 (m, 9H), 1.15 (s, 6H).

b) 9-bromo-2,2-dimethylnonan-1-ol

9-bromo-2,2-dimethyl-nonanoic acid ethyl ester (Example 103, step a) (1.27g) in dry diethyl ether solution (40 mL) at 0 ℃ in the N ₂ was added dropwise to the hydride diisobutyl Butyl aluminum (1 M in toluene, 9.5 mL). The reaction mixture was stirred at 0&lt;0&gt;C for 1.25 h then quenched (150 mL). The mixture was stirred for 15 min then EtOAc (EtOAc)EtOAc. Purification by gel chromatography eluting with 0 to 25% ethyl acetate in cyclohexane afforded the subtitle compound as a colourless liquid. The yield was 0.56 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 3.41 (t, J = 6.9 Hz, 2H), 3.31 (d, J = 5.2 Hz, 2H), 1.91-1.79 (m, 2H), 1.47-1.39 (m, 2H), 1.34-1.21 (m, 8H), 0.86 (s, 6H). An unobserved exchangeable proton.

c) (9-(9-Hydroxy-8,8-dimethylindolyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-iso Propylthiazole-4-yl)methanone

(2-Isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (0.5 g) was dissolved in methanol and applied to an SCX column pre-wetted with methanol. The column was washed with methanol and dissolved in 2M ammonia in methanol. The eluent evaporates in vacuo to yield the material as a free base (0.26 g). Addition to 9-bromo-2,2-dimethylnonan-1-ol in acetonitrile (10 mL) and triethylamine (0.23 mL) (Example 103, step b) (0.25 g) The resulting mixture was heated at 60 ° C for 15 h. The volatiles were evaporated in vacuo and the title compound was purified eluting eluting eluting eluting eluting eluting eluting Dissolved in 10% methanol to give the subtitle compound as an off-white solid. The yield was 0.37 g.

m/z 480(M+H) ⁺

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.87 (s, 1H), 4.08 - 3.91 (m, 1H), 3.80 - 3.75 (m, 3H), 3.45 - 3.32 (m, 2H), 3.35 - 3.30 (m) , 2H), 3.06-2.84 (m, 3H), 2.48-2.31 (m, 2H), 2.17-2.03 (m, 2H), 1.96-1.83 (m, 2H), 1.66-1.51 (m, 4H), 1.43 (d, J = 6.9 Hz, 6H), 1.39-1.31 (m, 4H), 1.30-1.16 (m, 6H), 0.86 (s, 6H). An unobserved exchangeable proton.

d) 9-(4-(2-Isopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)-2,2- Dimethylfurfural

Trifluoroacetic acid (0.059 mL) was added to (9-(9-hydroxy-8,8-dimethylindenyl)-1-oxa-4,9-diazaspiro[5.5] under argon at 0 °C. a solution of the undecane-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 103, step c) (0.37 g) in DCM (20 mL) Then, Dess-Martin periodinane (0.49 g) was added. The reaction mixture was stirred at rt EtOAc EtOAc (EtOAc)EtOAc. The phases were separated and the aqueous layer was extracted with ethyl acetate (×2). The combined organic solution was washed with a saturated sodium bicarbonate solution. Acetic acid (0.066 mL) was added to EtOAc (EtOAc)EtOAc. The yield was 0.40 g.

m/z 478(M+H) ⁺

_{^{1 H NMR (400MHz, CDCl 3}} ) δ9.46-9.41 (s, 1H), 7.87 (s, 1H), 4.05-3.89 (m, 3H), 3.82-3.67 (m, 5H), 3.36-3.26 (m , 3H), 2.87-2.79 (m, 4H), 2.09-2.02 (m, 2H), 1.77-1.65 (m, 2H), 1.47-1.36 (m, 10H), 1.32-1.13 (m, 6H), 1.03 (s, 6H).

e) (R)-4-hydroxy-7-(1-hydroxy-2-(9-(4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)-2,2-dimethylhydrazino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

Acetic acid (0.066 mL) was added to 9-(4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl )-2,2-dimethylfurfural

(Example 103, step d) (0.4 g) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride Salt (WO 2007027134, Example 1, step d) (0.3g) and The molecular sieves were in a mixture of anhydrous methanol (10 mL). The mixture was stirred for 5 min and then cooled to 0 °C. Sodium triethoxysulfonium borohydride (0.16 g) was added and the resulting mixture was stirred at RT for 16.5 h. The solution was filtered and concentrated in vacuo. The residue is dissolved in a mixture of acetonitrile and water and purified by preparative HPLC (Phenomenex , Gradient: 10 to 40% acetonitrile in 0.1% aqueous formic acid). The product-containing fractions were combined and lyophilized to give the title compound as a white solid. The yield was 0.10 g.

m/z 688(M+H) ⁺

¹ H NMR (400 MHz, D _{4 -} MeOH) δ 8.51 (s, 1H), 7.88 (s, 1H), 6.92 (d, J = 8.3 Hz, 1H), 6.70 (d, J = 8.3 Hz, 1H) , 4.93-4.88 (m, 1H), 3.92-3.53 (m, 6H), 3.32-3.25 (m, 1H), 3.11-2.52 (m, 10H), 1.98 (s, 2H), 1.82-1.48 (br m , 3H), 1.37 (d, J = 6.9 Hz, 6H), 1.38-1.31 (m, 5H), 1.30-1.22 (m, 6H), 0.91 (s, 6H). Five unobserved exchangeable protons.

Example 104

(R)-4-mercapto-7-(1-hydroxy-2-(4-(2-(4-(5-methylthiophen-3-carbonyl)-1-oxa-4,9-diaza) Heterospiro[5.5]undecane-9-yl)ethoxy)benzylamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

a) 4-(5-methylthiophene-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester

Adding O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.91 g) to 1-oxa-4,9 -diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester hydrochloride (WuXi PharmaTech) (0.5 g), 5-methyl-thiophene-3-carboxylic acid (0.243 g) and triethylamine (0.95 mL) in a solution in DMF (10 mL). The reaction mixture was poured into brine (40 mL) The combined organic solution was washed with water (150 mL) brine brine. Purification by gel chromatography eluting with 0 to 10% methanol in dichloromethane afforded the subtitle compound. The yield was 0.34 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.23 (s, 1H), 6.81 (s, 1H), 3.72 (s, 6H), 3.46 (s, 3H), 3.15 (s, 3H), 2.49 (d, J = 1.1 Hz, 3H), 1.82 (br d, J = 13.5 Hz, 2H), 1.45 (s, 9H).

b) (5-methylthiophen-3-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone

Adding trifluoroacetic acid (2.5 mL) to 4-(5-methylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid Butyl ester

(Example 104, step a) (0.34 g) in EtOAc (EtOAc)EtOAc. The solvent was evaporated in vacuo. Toluene (3 x 25 mL) was added and the mixture was evaporated in vacuo. The residue was dissolved in methanol (20 mL) and applied to EtOAc EtOAc. The column was washed with methanol (40 mL) and EtOAc (EtOAc) The eluent was evaporated in vacuo to give the subtitle compound as a colourless oil. The yield is 0.25 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ7.50 (s, 1H), 6.88 (s, 1H), 3.67-3.57 (m, 2H), 3.49 (s, 2H), 3.38 (s, 4H), 2.68 (s, 2H), 2.45 (d, J = 1.1 Hz, 3H), 1.57 (s, 2H), 1.37 (s, 2H). An unobserved exchangeable proton.

c) (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl (4-(2-(4) -(5-methylthiophene-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethoxy)benzyl)carbamic acid third Butyl ester

Triethylamine (0.25 mL) was added to (5-methylthiophen-3-yl)(1-oxa-4,9-diazaspiro[5.5]undec-4-yl)methanone ( Example 104, step b) (0.195 g) in EtOAc (2.5 mL). This solution is added to (R)-methanesulfonic acid 2-(4-((T-butoxycarbonyl(2-hydroxy-2-(4-hydroxy-2-yloxy-2,3-dihydrobenzo) [d] Thiazol-7-yl)ethyl)amino)methyl)phenoxy)ethyl ester (Example 108, step c) (0.5 g) in acetonitrile (2.5 mL). The resulting mixture was heated at 80 ° C for 48 hours. The reaction mixture was evaporated in vacuo. Purification by gel chromatography eluting with 0 to 10% methanol in dichloromethane afforded the title compound as a white solid. The yield was 0.33 g.

¹ H NMR (300 MHz, D _{4 -} MeOH) δ 7.42 (s, 1H), 7.07 (d, J = 6.7 Hz, 2H), 6.85 (d, J = 13.0 Hz, 4H), 6.70 (d, J = 8.2 Hz, 1H), 4.28 (s, 1H), 4.12 (s, 2H), 3.71 (s, 2H), 3.66-3.48 (m, 5H), 3.38-3.29 (m, 5H), 2.90 (s, 2H) , 2.80-2.55 (m, 4H), 2.48 (s, 3H), 1.94-1.89 (m, 1H), 1.44 (s, 4H), 1.37 (s, 6H). Three unobserved exchangeable protons.

d) (R)-4-hydroxy-7-(1-hydroxy-2-(4-(2-(4-(5-methylthiophene-3-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)ethoxy)benzylamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

Adding trifluoroacetic acid (1.2 mL) to (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl (4-(2-(5-methylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethoxy) A solution of benzyl) carbamic acid tert-butyl ester (Example 104, step c) (0.33 g) in DCM (5 mL). Toluene (20 mL) was added and the mixture was evaporated in vacuo. The residue was azeotroped with toluene (3 x 20 mL). Purified by preparative HPLC (Phenomenex , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The product-containing fractions were combined and lyophilized to give the title compound as a white solid. The yield was 0.12 g.

m/z 639(M+H) ⁺

¹ H NMR (400 MHz, D ₆ -DMSO) δ 8.13 (s, 1H), 7.45 (s, 1H), 7.22 (d, J = 8.34 Hz, 2H), 6.88-6.78 (m, 4H), 6.66 ( d, J = 8.27 Hz, 1H), 4.65 (m, 1H), 4.09-4.00 (m, 2H), 3.85-3.77 (m, 3H), 3.67-3.60 (m, 4H), 3.56-3.47 (m, 2H), 2.81-2.63 (m, 5H), 2.49-2.36 (m, 5H), 1.77-1.67 (m, 2H), 1.59-1.37 (m, 2H). Five unobserved exchangeable protons.

Example 105

(R)-4-mercapto-7-(1-hydroxy-2-(9-(4-(5-methylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)decylamino)ethyl)benzene well [d]thiazole-2(3H)-ketoformate

a) (9-(9-Hydroxymercapto)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(5-methylthiophen-3-yl)- ketone

Add a solution of 9-bromo-1-nonanol (0.335 g) in acetonitrile (3 mL) to (5-methylthiophen-3-yl)(1-oxa-4,9-diazaspiro[5.5 And a solution of triethylamine (0.278 g) in acetonitrile (7 mL). The resulting mixture was stirred overnight at 60 °C. The reaction mixture was evaporated with EtOAc EtOAc m. Purification by silica gel chromatography eluting with 0 to 10% methanol in dichloromethane afforded the subtitle compound as a white solid. The yield was 0.3 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 7.54 (s, 1H), 6.91 (s, 1H), 4.32 (t, J = 5.1 Hz, 1H), 3.66 (s, 2H), 3.52 (s, 2H), 3.41-3.33 (m, 3H), 2.95 (s, 4H), 2.45 (d, J = 1.1 Hz, 3H), 2.02 (s, 3H), 1.64 (s, 4H), 1.40 (t, J) = 6.8 Hz, 2H), 1.26 (s, 11H). An unobserved exchangeable proton.

b) (R)-4-hydroxy-7-(1-hydroxy-2-(9-(4-(5-methylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)nonylamino)ethyl)benzo[d]thiazole-2(3H)-one formate

Trifluoroacetic acid (0.055 mL) was added to (9-(9-hydroxyindenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4- at 0 °C under argon. (5-Methylthiophen-3-yl)methanone (Example 105, step a) (0.3 g) in EtOAc. The resulting mixture was stirred for 5 min and then Dess-Martin periodinane (0.45 g) was added. The mixture was stirred at room temperature for 1 h then EtOAc EtOAc m. The layers were separated and the aqueous layer was extracted with ethyl acetate (2×40mL). Acetic acid (0.159 mL) was added to a combined organic layer dried over magnesium sulfate, filtered and evaporated in vacuo. The yellow oil was dissolved in dry methanol (15 mL). Additive R)-7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) 0.28g), followed by acetic acid (0.061mL) and Molecular sieves. After stirring at room temperature for 5 min, the reaction mixture was cooled to 0 °C. Sodium triethoxysulfonium borohydride (0.15 g) was added and the mixture was stirred at room temperature for 18 h. The reaction mixture was filtered and the solvent was evaporated in vacuo. The residue was purified by preparative HPLC (Phenomenex , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The product-containing fractions were combined and lyophilized to give the title compound as a white solid. The yield was 0.07 g.

m/z 631(M+H) ⁺

^{1 H NMR (400MHz, D6-} DMSO) δ8.24 (s, 1H), 7.45 (s, 1H), 6.85-6.80 (m, 2H), 6.67 (d, J = 8.3Hz, 1H), 4.67 (t , J = 6.5 Hz, 1H), 3.63 - 3.21 (m, 6H), 2.75 (d, J = 6.5 Hz, 2H), 2.63 (t, J = 7.3 Hz, 2H), 2.40 (d, J = 1.1 Hz) , 3H), 2.38-2.16 (m, 6H), 1.70-1.61 (m, 2H), 1.47-1.29 (m, 6H), 1.19 (s, 10H). Five unobserved exchangeable protons.

Example 106

(R)-7-(2-(4-(2-(4-(benzo[d]thiazole-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecene Alkan-9-yl)ethoxy)benzylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-onecarboxylate

a) 4-(benzo[d]thiazole-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester

Adding O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1.45 g) to 1-oxa-4,9 -Diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester hydrochloride (WuXi PharmaTech) (0.8 g), 1,3-benzothiazole-2-carboxylic acid (0.588 g) and triethyl A solution of the amine (1.51 mL) in EtOAc m. The DMF volume was reduced in vacuo and the reaction mixture was partitioned between water (30mL) and ethyl acetate (30mL). The organic layer was washed with EtOAc EtOAc m. Purification by silica gel chromatography eluting with 0 to 10% methanol in dichloromethane to give the subtitle compound as a yellow oil. The yield was 0.83 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 8.23 - 8.09 (m, 2H), 7.63 - 7.52 (m, 2H), 4.34 (s, 1H), 4.20 (s, 1H), 3.78 (s, 2H) ), 3.71 (s, 1H), 3.64 - 3.51 (m, 3H), 3.08 (s, 2H), 1.76 (d, J = 13.7 Hz, 2H), 1.55-1.34 (m, 11H).

b) Benzene [d]thiazol-2-yl (1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone

Adding trifluoroacetic acid (5 mL) to 4-(benzo[d]thiazole-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid Butyl ester (Example 106, step a) (0.83 g) in EtOAc (EtOAc) The solvent was evaporated in vacuo. The residue was dissolved in DCM:methanol (1:1) (5 mL) and applied to EtOAc (EtOAc) The column was washed with methanol (60 mL) and EtOAc (EtOAc) The eluent was evaporated in vacuo to give the subtitle compound as a colourless oil. The yield was 0.57 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 8.18-8.13 (m, 1H), 8.10-8.07 (m, 1H), 7.60-7.50 (m, 2H), 4.25 (t, J = 4.8 Hz, 1H) ), 4.17 (s, 1H), 4.04 (s, 1H), 3.70 (s, 2H), 3.65 (d, J = 5.5 Hz, 1H), 3.56 (s, 1H), 2.73 - 2.46 (m, 4H) , 1.62-1.54 (m, 2H), 1.50-1.36 (m, 2H).

c) (R)-4-(2-(4-(benzo[d]thiazole-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9- Ethyl)benzyl (2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl)aminocarbamic acid Tributyl ester

Addition of triethylamine (0.25 mL) to benzo[d]thiazol-2-yl(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone (implementation) Example 106, step b) (0.22 g) in EtOAc (EtOAc) This solution is added to (R)-methanesulfonic acid 2-(4-((T-butoxycarbonyl(2-hydroxy-2-(4-hydroxy-2-yloxy-2,3-dihydrobenzo) [d] Thiazol-7-yl)ethyl)amino)methyl)phenoxy)ethyl ester (Example 108, step c) (0.5 g) in acetonitrile (2.5 mL). The resulting mixture was heated at 80 ° C for 48 hours. The reaction mixture was evaporated in vacuo. Purification by silica gel chromatography, eluting with 0 to 10% methanol in dichloromethane afforded the sub-title compound as a creamy white foam. The yield was 0.34 g.

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ11.49 (s, 1H), 9.93 (s, 1H), 8.21 (d, J = 7.5Hz, 1H), 8.14 (d, J = 7.7Hz, 1H) , 7.64 - 7.56 (m, 2H), 7.07 (s, 2H), 6.93 - 6.79 (m, 2H), 6.78-6.67 (m, 2H), 5.68 (s, 1H), 4.73 (s, 1H), 4.35 -4.22 (m, 3H), 4.07 (s, 4H), 3.76 (s, 2H), 3.70 (s, 1H), 3.62 (s, 1H), 3.17 (s, 6H), 2.67 (s, 2H), 1.77 (s, 1H), 1.62 (s, 1H), 1.35 (s, 3H), 1.27 (s, 6H). An unobserved exchangeable proton.

d) (R)-7-(2-(4-(2-(4-(benzo[d]thiazole-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5] Monoalkane-9-yl)ethoxy)benzylamino)-1-hydroxyethyl)-4-hydroxybenzene well [d]thiazole-2(3H)-one formate

Adding trifluoroacetic acid (1.25 mL) to (R)-4-(2-(4-(benzo[d]thiazol-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)ethoxy)benzyl (2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazole-7-yl) Ethyl) butyl methacrylate (Example 106, step c) (0.33 g) in DCM (4 mL) Toluene (20 mL) was added and the mixture was evaporated in vacuo. The residue was azeotroped with toluene (3 x 20 mL). Purified by preparative HPLC (Phenomenex , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The product-containing fractions were combined and lyophilized to give the title compound as a white solid. The yield was 0.096 g.

m/z 676(M+H) ⁺

¹ H NMR (400 MHz, D ₄ -MeOH) δ 8.44 (s, 1H); 8.09-8.00 (m, 2H); 7.58-7.48 (m, 2H); 7.37 (t, J = 6.6 Hz, 2H); 7.00 (t, J = 9.4 Hz, 2H); 6.91 (d, J = 8.3 Hz, 1H); 6.71 (d, J = 8.3 Hz, 1H); 4.93-4.88 (m, 1H); 4.43 - 3.38 (m , 1H); 4.37-4.34 (s, 1H); 4.27-4.20 (m, 2H); 4.14 (s, 2H); 3.87-3.82 (m, 2H); 3.81-3.76 (m, 1H); 3.71 (s , 1H); 3.20-3.11 (m, 2H); 3.12-3.01 (m, 4H); 2.96-2.82 (m, 2H); 2.10-2.01 (m, 2H); 1.88-1.76 (m, 2H). Five unobserved exchangeable protons.

Example 107

(R)-7-(2-(9-(4-(benzo[d]thiazol-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)decylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-onecarboxylate

a) benzo[d]thiazol-2-yl(9-(9-hydroxyindolyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone

Add a solution of 9-bromo-1-nonanol (0.335 g) in acetonitrile (3 mL) to benzo[d]thiazol-2-yl (1-oxa-4,9-diazaspiro[5.5] A solution of the undecane-4-yl)methanone (Example 106, step b) (0.34 g) and triethylamine (0.278 mL) in acetonitrile (7 mL). The resulting mixture was stirred overnight at 60 °C. The reaction mixture was evaporated with EtOAc EtOAc m. Purification by silica gel chromatography eluting with 0 to 10% methanol in dichloromethane afforded the subtitle compound as a white solid. The yield was 0.34 g.

¹ H NMR (400 MHz, D ₆ -DMSO) δ 8.22 (d, J = 7.7 Hz, 1H), 8.14 (d, J = 7.9 Hz, 1H), 7.65-7.56 (m, 2H), 4.37 (s, 1H), 4.31 (t, J = 5.1 Hz, 1H), 4.27 (s, 1H), 3.80 (s, 2H), 3.72 (s, 1H), 3.63 (s, 1H), 3.41-3.33 (m, 2H) ), 2.99 (s, 4H), 2.10 (s, 2H), 1.86 (s, 2H), 1.64 (s, 2H), 1.40 (s, 3H), 1.27 (s, 11H).

b) (R)-7-(2-(9-(4-(benzo[d]thiazole-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane -9-yl)decylamino)-1-hydroxyethyl)-4-hydroxybenzo [d]thiazole-2(3H)-ketoformate

Trifluoroacetic acid (0.056 mL) was added to benzo[d]thiazol-2-yl(9-(9-hydroxyindenyl)-1-oxa-4,9-diaza snail at 0 ° C under argon. [5.5] undecane-4-yl)methanone (Example 107, step a) (0.33 g) in EtOAc. The resulting mixture was stirred for 5 min and then Dess-Martin periodinane (0.46 g) was added. The mixture was stirred at room temperature for 1 h. Saturated sodium thiosulfate solution (14 mL), sat. sodium bicarbonate (14 mL) and EtOAc (30 mL). The layers were separated and the aqueous extracted with EtOAc EtOAc. Acetic acid (0.159 mL) was added to a combined organic layer dried over magnesium sulfate, filtered and evaporated in vacuo. The yellow oil was dissolved in dry MeOH (15 mL). ( R )-7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.28g) followed by acetic acid (0.062mL) and 3 Molecular sieves. After stirring at room temperature for 5 min, the reaction mixture was cooled to 0 °C. Sodium triethoxysulfonium borohydride (0.15 g) was added and the mixture was stirred at room temperature for 18 h. The reaction mixture was filtered and the solvent was evaporated in vacuo. The residue was purified by preparative HPLC (Phenomenex , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The product-containing fractions were combined and lyophilized to give the subtitle compound as a white solid. The yield was 0.14 g.

m/z 668(M+H) ⁺

_{^{1 H NMR (400MHz, D 6}} -DMSO) δ8.23 (s, 1H), 8.17-8.14 (m, 1H), 8.09 (d, J = 7.9Hz, 1H), 7.60-7.49 (m, 2H), 6.84 (d, J = 8.3 Hz, 1H), 6.68 (d, J = 8.29 Hz, 1H), 4.74 - 4.64 (m, 1H), 4.29 - 4.24 (m, 1H), 4.20 (s, 1H), 3.72 -3.62(m,3H),3.56(s,1H), 2.79(d,J=5.5Hz,2H),2.69-2.62(m,2H),2.42-2.13(m,6H),1.74-1.64(m , 2H), 1.63-1.30 (m, 6H), 1.19 (d, J = 9.6 Hz, 10H). Five unobserved exchangeable protons.

Example 108

(R)-4-hydroxy-7-(1-mercapto-2-(4-(2-(4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)ethoxy)benzylamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

a) 4-(2-mercaptoethoxy)benzaldehyde

2-Bromoethanol (5.8 mL) and potassium carbonate (11.3 g) were continuously added to a solution of 4-hydroxybenzaldehyde (5.0 g) in MeCN (100 mL). The resulting mixture was stirred and heated under reflux for 7 days under argon. The reaction mixture was cooled to rt EtOAc (EtOAc)EtOAc. The organic layer was washed with brine (100 mL). The aqueous layer was combined and extracted with EtOAc (50 mL). The combined organic layer was dried with sodium sulfate, filtered and evaporated in vacuo. The yield was 6.4 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ9.90 (s, 1H), 7.87-7.81 (m, 2H), 7.06-6.98 (m, 2H), 4.18 (dd, J = 5.0,4.0Hz, 2H), 4.04-3.97 (m, 2H), 2.18-1.96 (m, 1H).

b) 2-(4-Mexylphenoxy)ethyl methanesulfonate

Methanesulfonium chloride (3.3 mL) was added dropwise at 0 ° C under argon over 5 min to 4-(2-hydroxyethoxy)benzaldehyde (Example 108, Step a) (6.4 g) and triethylamine (6.4) mL) in a solution in DCM (30 mL). The resulting mixture was stirred at 0 °C for 10 min and then with RT for 1 h. The reaction mixture was washed with brine (2×30 mL) The residue was purified by EtOAc (EtOAc) elut elut elut elut elut The yield was 8.87 g. ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.92-9.87 (m, 1H), 7.89-7.81 (m, 2H), 7.05-6.98 (m, 2H), 4.62-4.58 (m, 2H), 4.36-4.32 (m, 2H), 3.10 (s, 3H).

c) (R)-Methanesulfonic acid 2-(4-((T-butoxycarbonyl(2-hydroxy-2-(4-hydroxy-2-yloxy-2,3-dihydrobenzo[d] Thiazol-7-yl)ethyl)amino)methyl)phenoxy)ethyl ester

2-(4-Mexylphenoxy)ethyl methanesulfonate (Example 108, step b) (1.7 g), ( R )-7-(2-amino-1-hydroxyethyl)-4 a mixture of -hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (2.0 g) and acetic acid (0.44 mL) in anhydrous DMF (40 mL) In; 4 The mixture was stirred for 15 min in the presence of molecular sieves. The reaction mixture was cooled to 0 ° C and sodium triethyl sulfoxyborohydride (2.2 g) was added in portions at intervals of 2 min. The resulting mixture was stirred at RT for 1 h. Di-tert-butyl dicarbonate (1.8 g) was added to the reaction mixture and stirred for 2 h. Saturated sodium bicarbonate solution (100 mL) and EtOAc (100 mL). The layers were separated and the aqueous layer was extracted with EtOAc (× 4). The combined organic layers were dried over sodium sulfate, filtered and evaporated in vacuo. The residue was purified by EtOAc EtOAc (EtOAc) The yield was 2.25 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 7.05 (d, J = 7.6 Hz, 2H), 6.80 (d, J = 8.0 Hz, 3H), 6.70 (d, J = 8.0 Hz, 1H), 4.81 (bs) , 1H), 4.54 (t, J = 4.3 Hz, 2H), 4.34 - 4.26 (m, 1H), 4.24 - 4.08 (m, 3H), 3.57 - 3.42 (m, 1H), 3.33 - 3.23 (m, 1H) ), 3.07 (s, 3H), 1.49 (s, 9H). Three unobserved exchangeable protons.

d) (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl (4-(2-(4) -(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diaza Spiral [5.5]undecane-9-yl)ethoxy)benzyl)carbamic acid tert-butyl ester

(R)-Methanesulfonic acid 2-(4-((T-butoxycarbonyl(2-hydroxy-2-(4-hydroxy-2-yloxy)-2,3-dihydrobenzo[d]thiazole a solution of -7-yl)ethyl)amino)methyl)phenoxy)ethyl ester (Example 108, step c) (0.295 g) in MeCN (1.1 mL). (2-Isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 22, step b) (0.150 g) and a solution of triethylamine (0.11 mL) in MeCN (1.1 mL). The resulting mixture was stirred at 60 ° C under argon for 34 h. After cooling to RT, the solvent was removed in vacuo. The residue was taken up in EtOAc (EtOAc)EtOAc. The yellow solid was purified by EtOAc (EtOAc) elute The yield was 0.135 g.

m/z 768(M+H) ⁺

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.86 (s, 1H), 6.83 (br s, 2H), 6.71-6.46 (m, 4H), 4.78 (br s, 1H), 4.46-2.51 (m, 19H ), 1.90 (br s, 4H), 1.53 (s, 9H), 1.39 (s, 3H), 1.29 (s, 3H). Three unobserved exchangeable protons.

e) (R)-4-hydroxy-7-(1-hydroxy-2-(4-(2-(4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-) Diazaspiro[5.5]undecane-9-yl)ethoxy)benzyl Amino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

Adding trifluoroacetic acid (0.7 mL) to (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl (4-(2-(4-(2-Isopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethoxy) Benzyl) carbamic acid tert-butyl ester (Example 108, step d) (0.122 g) in DCM ( 1.5 mL). The resulting mixture was stirred at RT for 10 min. Toluene (15 mL) was added and the reaction mixture was evaporated in vacuo. The residue was azeotroped twice with MeCN. Viscous yellow residue purified by HPLC (Phenomenex Gemini) , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The product-containing fractions were combined and lyophilized to give the title compound as a white solid. The yield was 0.052 g.

m/z 668(M+H) ⁺

¹ H NMR (400 MHz, D _{4 -} MeOH) δ 8.47 (s, 1H), 7.78 (s, 1H), 7.36 (d, J = 8.19 Hz, 2H), 6.98 (s, 2H), 6.91 (d, J = 8.36 Hz, 1H), 6.71 (d, J = 8.30 Hz, 1H), 4.90 (dd, J = 9.4, 4.1 Hz, 1H), 4.18 (s, 2H), 4.11 (s, 2H), 3.75 ( s, 5H), 3.63 (s, 1H), 3.40 (m, 1H), 3.19-2.80 (m, 8H), 1.95 (s, 2H), 1.75 (s, 1H), 1.65 (s, 1H), 1.38 (d, J = 6.9 Hz, 6H). Five unobserved exchangeable protons.

Example 109

(R)-4-hydroxy-7-(1-hydroxy-2-(9-(4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)decylamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

a) (9-(9-Hydroxymercapto)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-isopropylthiazol-4-yl) Ketone

A solution of 9-bromo-1-nonanol (0.316 g) in MeCN (3 mL) was added to (2-isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[ 5.5] Idecan-4-yl)methanone trifluoroacetate (Example 22, step b) (0.40 g) and triethylamine (0.395 mL) in MeCN (7 mL). The resulting mixture was stirred at 60 ° C under argon for 16 h. After cooling to RT, the solvent was removed in vacuo. The residue was taken up in EtOAc (EtOAc)EtOAc. The orange solid was purified by EtOAc (EtOAc) elute The yield was 0.325 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.87 (s, 1H), 3.97 (s, 2H), 3.76 (s, 3H), 3.63 (t, J = 6.6Hz, 2H), 3.36 (s, 3H) , 2.92 (s, 4H), 2.25 (s, 2H), 2.14 - 2.03 (m, 2H), 1.84 (s, 2H), 1.63 (s, 2H), 1.60-1.50 (m, 2H), 1.42 (d , J = 6.8 Hz, 6H), 1.42-1.21 (m, 10H).

b (R)-4-hydroxy-7-(1-hydroxy-2-(9-(4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)nonylamino)ethyl)benzo[d]thiazole-2(3H)-one formate

Adding trifluoroacetic acid (0.027 mL) to (9-(9-hydroxyindenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl) (2-iso Propylthiazole-4-yl)methanone (Example 109, step a) (0.16 g) in DCM (EtOAc. The resulting mixture was stirred for 5 min and then Dess-Martin periodinane (0.225 g) was added. The mixture was stirred at RT for 45 min. A saturated solution of sodium thiosulfate (6 mL), EtOAc (EtOAc) (EtOAc) The layers were separated and the aqueous extracted with EtOAc EtOAc. The combined organic layers were washed with EtOAc EtOAc EtOAc. The viscous orange oil was dissolved in anhydrous MeOH (6.6 mL) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one salt was added. Acid salt (WO 2007027134, Example 1, step d) (0.138 g), followed by acetic acid (0.03 mL) and Molecular sieves. After stirring at RT for 5 min, the reaction mixture was cooled to 0 &lt;0&gt;C and sodium triacetoxyborohydride (0.075 g) The mixture was stirred at RT for 1 h then a further portion of sodium tris-sodium succinate (0.075 g). The reaction mixture was filtered and the solvent was evaporated in vacuo. The residue was purified by HPLC (Phenomenex Gemini) , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The product-containing fractions were combined and lyophilized to give the title compound as a white solid. The yield was 0.060 g.

m/z 660(M+H) ⁺

¹ H NMR (400 MHz, D _{4 -} MeOH) δ 8.52 (s, 1H), 7.86 (s, 1H), 6.94 (d, J = 8.4 Hz, 1H), 6.52 (d, J = 8.3 Hz, 1H) , 4.98-4.89 (m, 1H), 3.90-3.58 (m, 6H), 3.37-3.25 (m, 1H), 3.15-3.02 (m, 3H), 3.01-2.93 (m, 3H), 2.91-2.65 ( m, 4H), 2.10 - 1.96 (m, 2H), 1.83-1.54 (m, 6H), 1.41-1.28 (m, 16H). Five unobserved exchangeable protons.

Example 110

(R)-4-hydroxy-7-(1-hydroxy-2-(4-(2-(4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)ethoxy)benzylamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

a) (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl (4-(2-(4) -(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro [5.5] Decadin-9-yl)ethoxy)benzyl)carbamic acid tert-butyl ester

(R)-Methanesulfonic acid 2-(4-((T-butoxycarbonyl(2-hydroxy-2-(4-hydroxy-2-yloxy)-2,3-dihydrobenzo[d]thiazole a solution of -7-yl)ethyl)amino)methyl)phenoxy)ethyl ester (Example 108, step c) (0.426 g) in MeCN (2 mL). 5-methylthiophen-2-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone trifluoroacetate (Example 9, step b) (0.154 g) and a solution of triethylamine (0.15 mL) in MeCN (5 mL). The resulting mixture was stirred at 60 ° C under argon for 16 h and then at 80 ° C for an additional 3 h. After cooling to RT, the solvent was removed in vacuo. The residue was taken up in EtOAc (EtOAc)EtOAc. The brown solid was purified by EtOAc (EtOAc) elute The yield was 0.246 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.11 (d, J = 3.6Hz, 1H), 6.89 (s, 2H), 6.70-6.59 (m, 4H), 6.51 (d, J = 8.2Hz, 1H) , 4.76 (s, 1H), 4.20-4.10 (m, 4H), 3.74 (s, 4H), 3.59-3.44 (m, 4H), 2.93-2.77 (m, 4H), 2.61 (s, 2H), 2.47 (s, 3H), 1.93-1.84 (m, 2H), 1.74-1.60 (m, 2H), 1.52 (s, 9H). Three unobserved exchangeable protons.

b) (R)-4-hydroxy-7-(1-hydroxy-2-(4-(2-(4-(5-methylthiophene-2-carbonyl)-) 1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethoxy)benzylamino)ethyl)benzo[d]thiazole-2(3H)-one Formate

Adding trifluoroacetic acid (0.75 mL) to (R)-methanesulfonic acid 2-(4-((T-butoxycarbonyl)-(2-hydroxy-2-(4-hydroxy-2-yloxy-2, 3-Dihydrobenzo[d]thiazol-7-yl)ethyl)amino)methyl)phenoxy)ethyl ester (Example 110, step a) (0.243 g) in DCM (3 mL) in. The resulting mixture was stirred at RT for 10 min. Toluene (30 mL) was added and the mixture was evaporated in vacuo (×3). The residue was purified by HPLC (Phenomenex , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The product-containing fractions were combined and lyophilized to give the title compound as a white solid. The yield was 0.65 g.

m/z 639(M+H) ⁺

¹ H NMR (400 MHz, D ₄ -MeOH) δ 8.44 (s, 1H), 7.40-7.35 (m, 2H), 7.19 (d, J = 3.7 Hz, 1H), 7.02-6.97 (m, 2H), 6.92 (d, J = 8.36 Hz, 1H), 6.78-6..76 (m, 1H), 6.71 (d, J = 8.30 Hz, 1H), 4.93-4.88 (m, 1H), 4.21 (t, J) =5.17Hz,2H),4.13(s,2H), 3.76-3.70(m,4H),3.61(s,2H),3.11-3.07(m,2H),3.07-3.01(m,2H),3.03- 2.91 (m, 2H), 2.88-2.76 (m, 2H), 2.47 (d, J = 1.0 Hz, 3H), 2.02-1.93 (m, 2H), 1.74-1.61 (m, 2H). Five unobserved exchangeable protons.

Example 111 ( R ) 4-hydroxy-7-(1-hydroxy-2-(9-(4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro[5.5] eleven Cycloalkane-9-yl)decylamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

a) (9-(9-Hydroxymercapto)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(5-methylthien-2-yl)- ketone

A solution of 9-bromo-1-nonanol (0.314 g) in MeCN (3 mL) was added to (5-methylthiophen-2-yl)(1-oxa-4,9-diazaspiro[5.5 a solution of the undecane-4-yl)methanone trifluoroacetate (Example 9, step b) (0.263 g) and triethylamine (0.261 mL) in MeCN (7 mL). The resulting mixture was stirred at 60 ° C under argon for 16 h. After cooling to RT, the solvent was removed in vacuo. The residue was taken up in EtOAc (EtOAc)EtOAc. The white solid was purified by EtOAc (EtOAc) elute The yield was 0.352 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.10 (d, J = 3.6Hz, 1H), 6.71 (dd, J = 3.6,1.2Hz, 1H), 3.73 (s, 4H), 3.66-3.60 (m, 4H), 3.22 (s, 2H), 2.84 (s, 3H), 2.50 (d, J = 1.0 Hz, 3H), 2.15 (s, 1H), 2.11-1.97 (m, 2H), 1.83 (s, 2H) ), 1.61-1.50 (m, 3H), 1.32 (s, 11H). An unobserved exchangeable proton.

b) (R)-4-hydroxy-7-(1-hydroxy-2-(9-(4-(5-methylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)nonylamino)ethyl)benzo[d]thiazole-2(3H)-one formate

Adding trifluoroacetic acid (0.064 mL) to (9-(9-hydroxyindenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl) (5-A) Thiophen-2-yl)methanone (Example 111, step a) (0.350 g) in DCM (15 mL) EtOAc. The resulting mixture was stirred for 5 min and then Dess-Martin periodinane (0.527 g) was added. The mixture was stirred at RT for 1 h. Saturated sodium thiosulfate solution (14 mL), sat. sodium bicarbonate (14 mL) and EtOAc (30 mL). The layers were separated and the aqueous extracted with EtOAc EtOAc. The combined organic layers were washed with EtOAc EtOAc EtOAc. Yellow oil was dissolved in anhydrous MeOH (15.6 mL) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride Salt (WO 2007027134, Example 1, step d) (0.327 g), followed by acetic acid (0.071 mL) and Molecular sieves. After stirring for 5 min at RT, the reaction mixture was cooled to EtOAc, EtOAc (EtOAc) Another portion of sodium triethoxysulfonate hydride (0.176 g) was added and the mixture was stirred at RT for 17 h. The reaction mixture was filtered and the solvent was evaporated in vacuo. The residue was purified by HPLC (Phenomenex , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The product-containing fractions were combined and lyophilized to give the title compound as a white solid. The yield was 0.127 g.

m/z 631(M+H) ⁺

_{^{1 H NMR (400MHz, D 4}} -MeOH) δ8.49 (s, 1H), 7.20 (d, J = 3.7Hz, 1H), 6.95 (d, J = 8.3Hz, 1H), 6.79-6.76 (m, 1H), 6.73 (d, J = 8.3 Hz, 1H), 4.97-4.91 (m, 1H), 3.76-3.72 (m, 4H), 3.62 (s, 2H), 3.20-3.04 (m, 4H), 3.03 -2.93(m,3H), 2.95-2.84(m,3H), 2.47(d,J=1.0Hz,3H),2.11-2.01(m,2H),1.80-1.62(m,6H),1.40-1.31 (m, 10H). Five unobserved exchangeable protons.

Example 112

( R )-4-hydroxy-7-(1-hydroxy-2-(4-(2-(4-(2-phenylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)ethoxy)benzylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 4-(2-phenylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester

1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester hydrochloride (WuXi PharmaTech) (0.60 g) in DMF (11 mL) Amine (1.1 mL) followed by 2-phenyl-1,3-thiazole-4-carboxylic acid (0.421 g) followed by O-(7-azabenzotriazol-1-yl)-N hexafluorophosphate , N, N', N'-tetramethyluronium (1.09 g) treatment. The resulting mixture was stirred at RT for 7 h. Remove the DMF in a vacuum. The residue was taken into EtOAc (EtOAc)EtOAc. The brown oil was purified by EtOAc (EtOAc) elute The yield was 0.857 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ8.00 (s, 1H), 7.92 (s, 2H), 7.47 (d, J = 4.1Hz, 3H), 4.13-3.96 (m, 2H), 3.83 (s, 3H), 3.73 (s, 2H), 3.29-3.13 (m, 2H), 1.94-1.83 (m, 2H), 1.62-1.51 (m, 3H), 1.45 (s, 9H).

b) (2-Phenylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone

Trifluoroacetic acid (3.8 mL) was added to 4-(2-phenylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid Butyl ester (Example 112, step a) (0.85 mg) in DCM (15.2 mL). The resulting mixture was stirred at RT for 1 h. Toluene (30 mL) was added and the mixture was evaporated in vacuo. The yellow residue was dissolved in methanol and applied to an SCX column pre-wetted with MeOH. The column was washed with MeOH and taken up in 2M ammonia MeOH. The eluent was evaporated in vacuo to give the subtitle compound as a brown oil. The yield was 0.629 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 7.98 (s, 1H), 7.95-7.90 (m, 2H), 7.49-7.44 (m, 3H), 4.07 (br s, 1H), 3.99 (br s, 1H) , 3.88-3.79 (m, 3H), 3.70 (br s, 1H), 3.02-2.81 (m, 3H), 2.80-2.69 (m, 1H), 1.83 (d, J = 13.7 Hz, 2H), 1.70- 1.44 (m, 2H). An unobserved exchangeable proton.

c) (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl (4-(2-(4) -(2-phenylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethoxy)benzyl)carbamic acid third Butyl ester

(R)-Methanesulfonic acid 2-(4-((T-butoxycarbonyl(2-hydroxy-2-(4-hydroxy-2-yloxy)-2,3-dihydrobenzo[d]thiazole A solution of -7-yl)ethyl)amino)methyl)phenoxy)ethyl ester (Example 108, step c) (0.619 g) in MeCN (3 mL) was added dropwise at RT under argon ( 2-Phenylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecethane-4-yl)methanone (Example 112, step b) (0.274 g) And a solution of triethylamine (0.22 mL) in MeCN (7.3 mL). The resulting mixture was stirred at 80 ° C under argon for 16 h. After cooling to RT, the solvent was removed in vacuo. The residue was taken up in EtOAc (EtOAc m. The residue was purified by EtOAc EtOAc (EtOAc) The yield was 0.42 g.

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.04-7.95 (m, 1H), 7.94-7.79 (m, 2H), 7.50-7.41 (m, 2H), 7.40-7.28 (m, 1H), 7.02-6.81 (m, 2H), 6.70-6.37 (m, 3H), 4.84-4.70 (m, 1H), 4.25-3.92 (m, 6H), 3.87-3.75 (m, 3H), 3.74-3.58 (m, 2H) , 3.57-3.36 (m, 2H), 2.99-2.77 (m, 3H), 2.76-2.43 (m, 3H), 2.01-1.86 (m, 2H), 1.84-1.58 (m, 2H), 1.52 (s, 9H). Three unobserved exchangeable protons.

d) (R)-4-mercapto-7-(1-hydroxy-2-(4-(2-(4-(2-phenylthiazole-4-carbonyl)-1-oxa-4,9- Diazaspiro[5.5]undecane-9-yl)ethoxy)benzylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

Adding trifluoroacetic acid (1.2 mL) to (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl (4-(2-(4-(2-phenylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethoxy) Benzyl) carbamic acid tert-butyl ester (Example 112, step c) (0.415 g). The resulting mixture was stirred at RT for 15 min. Toluene (40 mL) was added and the reaction mixture was evaporated in vacuo. The residue was purified by HPLC (Phenomenex , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The product-containing fractions were combined and lyophilized to give the title compound as a white solid. The yield was 0.176 g.

m/z 702(M+H) ⁺

¹ H NMR (400 MHz, D ₄ -MeOH) δ 8.07 (s, 1H), 7.94 (br s, 2H), 7.51-7.39 (m, 5H), 7.06 (br s, 2H), 6.93 (d, J) = 8.4 Hz, 1H), 6.71 (d, J = 8.4 Hz, 1H), 4.92-4.88 (m, 1H), 4.38 (br s, 2H), 4.24 - 4.14 (m, 2H), 4.03-3.49 (m , 10H), 3.38-3.28 (m, 2H), 3.08-3.00 (m, 2H), 2.33-2.22 (m, 2H), 1.87 (br s, 2H). Six unobserved exchangeable protons.

Example 113

(R)-4-hydroxy-7-(1-hydroxy-2-(9-(4-(2-phenylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5 Eldecane-9-yl)decylamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

a) (9-(9-Hydroxymercapto)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-phenylthiazol-4-yl)- ketone

A solution of 9-bromo-1-nonanol (0.346 g) in MeCN (3 mL) was added to (2-phenylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5 a solution of the undecane-4-yl)methanone (Example 112, step b) (0.355 g) and triethylamine (0.288 mL) in MeCN (7 mL). The resulting mixture was stirred at 60 ° C under argon for 17 h. After cooling to RT, the solvent was removed in vacuo. The residue was taken up in EtOAc (EtOAc)EtOAc. The yellow oil was purified by EtOAc (EtOAc) elute The yield was 0.378 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ8.02 (s, 1H), 8.00-7.87 (m, 2H), 7.57 (s, 1H), 7.49-7.42 (m, 2H), 4.16-4.08 (m, 2H ), 3.81 (s, 4H), 3.72 (br s, 1H), 3.63 (t, J = 6.6 Hz, 2H), 3.33 - 2.64 (m, 6H), 2.25 - 2.04 (m, 3H), 1.79 (s) , 3H), 1.61-1.50 (m, 3H), 1.39-1.25 (m, 9H).

b)( R ) - 4-hydroxy-7-(1-hydroxy-2-(9-(4-(2-phenylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undeccarbon Alkan-9-yl)decylamino)ethyl)benzene well [d]thiazole-2(3H)-ketoformate

Adding trifluoroacetic acid (0.059 mL) to (9-(9-hydroxyindenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-benzene The thiothiazol-4-yl)methanone (Example 113, step a) (0.373 g) in DCM (14 mL) The resulting mixture was stirred for 5 min and then Dess-Martin periodinane (0.489 g) was added. The mixture was stirred at RT for 1 h. Saturated sodium thiosulfate solution (14 mL), sat. sodium bicarbonate (14 mL) and EtOAc (30 mL). The layers were separated and the aqueous extracted with EtOAc EtOAc. The combined organic layers were treated with EtOAc (EtOAc)EtOAc. Yellow oil was dissolved in anhydrous MeOH (15 mL) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.303 g), followed by acetic acid (0.066 mL) and Molecular sieves. After stirring for 5 min at RT, the reaction mixture was cooled to <RTI ID=0.0></RTI><RTIgt;</RTI><RTIgt; The reaction mixture was filtered and the solvent was evaporated in vacuo. The residue was purified by HPLC (Phenomenex , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The product-containing fractions were combined and lyophilized to give the title compound as a white solid. The yield was 0.118 g.

m/z 694(M+H) ⁺

¹ H NMR (400 MHz, D ₄ -MeOH) δ 8.31 (s, 1H), 7.97 (br s, 1H), 7.99-7.91 (m, 2H), 7.51-7.44 (m, 3H), 6.95 (d, J=8.4 Hz, 1H), 6.72 (d, J=8.4 Hz, 1H), 4.97-4.89 (m, 1H), 3.99-3.86 (m, 2H), 3.85-3.63 (m, 4H), 3.14-3.01 (m, 2H), 3.00-2.91 (m, 3H), 2.90-2.56 (m, 4H), 2.12-1.97 (m, 2H), 1.85-1.47 (m, 6H), 1.43-1.22 (m, 10H) . A proton that is obscured by solvents and five unobserved exchangeable protons.

Example 114

(R)-4-hydroxy-7-(1-hydroxy-2-(4-(2-(4-(2-isopropylthiazol-5-carbonyl)-1-oxa-4,9-diaza) Hetero[5.5]undecane-9-yl)ethoxy)benzylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 4-(2-isopropylthiazol-5-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester

1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid tert-butyl ester hydrochloride (WuXi PharmaTech) (0.694 g) in DMF (13 mL) Amine (1.3 mL) followed by 2-isopropylthiazole-5-carboxylic acid (Example 55, step b) (0.406 g), followed by O-(7-azabenzotriazole-1- hexafluorophosphate Base) -N,N,N',N'-tetramethyluronium (1.26 g). The resulting mixture was stirred at RT for 16 h. Remove the DMF in a vacuum. The residue was taken into EtOAc (EtOAc)EtOAc. The viscous orange oil was purified by chromatography on silica gel eluting with EtOAc (EtOAc) The yield was 0.765 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.80 (s, 1H), 3.75 (d, J = 3.2Hz, 6H), 3.55 (d, J = 4.3Hz, 2H), 3.36-3.28 (m, 1H) , 3.15 (s, 2H), 1.89-1.78 (m, 2H), 1.58 (s, 2H), 1.45 (s, 9H), 1.42 (d, J = 6.9 Hz, 6H).

b) (2-Isopropylthiazole-5-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone

Trifluoroacetic acid (3.9 mL) was added to 4-(2-isopropylthiazol-5-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-carboxylic acid Tributyl ester (Example 113, step a) (0.765 g) in DCM (14.8 mL). The resulting mixture was stirred at RT for 1 h. Toluene (30 mL) was added and the mixture was evaporated in vacuo. The yellow residue was dissolved in methanol and applied to an SCX column pre-wetted with MeOH. The column was washed with MeOH and taken up in 2M ammonia MeOH. The eluent was evaporated in vacuo to give the subtitle compound as a yellow oil. The yield was 0.552 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.81 (s, 1H), 3.79-3.71 (m, 4H), 3.64-3.44 (m, 2H), 3.36-3.26 (m, 1H), 2.96-2.87 (m , 2H), 2.85-2.73 (m, 2H), 1.86-1.75 (m, 2H), 1.59 (s, 2H), 1.42 (d, J = 6.9 Hz, 6H). An unobserved exchangeable proton.

c) (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl (4-(2-(4) -(2-isopropylthiazol-5-carbonyl)-1-oxa-4,9-diaza Spiral [5.5]undecane-9-yl)ethoxy)benzyl)carbamic acid tert-butyl ester

(R)-Methanesulfonic acid 2-(4-((T-butoxycarbonyl(2-hydroxy-2-(4-hydroxy-2-yloxy)-2,3-dihydrobenzo[d]thiazole A solution of -7-yl)ethyl)amino)methyl)phenoxy)ethyl ester (Example 108, step c) (0.575 g) in MeCN (3 mL). 2-isopropylthiazol-5-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone (Example 114, step b) (0.229 g And a solution of triethylamine (0.21 mL) in MeCN (7.3 mL). The resulting mixture was stirred at 80 ° C under argon for 16 h. After cooling to RT, the solvent was removed in vacuo. The residue was taken up in EtOAc EtOAc m. The residue was purified by EtOAc (EtOAc) elute The yield was 0.395 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ 7.69 (s, lH), 7.01-6.83 (s, 2H), 6.70-6.62 (m, 3H), 6.57-6.47 (m, 1H), 4.79 (s, 1H) , 4.25-4.06 (m, 3H), 3.77-3.63 (m, 4H), 3.57-3.39 (m, 4H), 3.34-3.26 (m, 2H), 2.90 (s, 4H), 2.61 (s, 2H) , 1.95-1.85 (m, 2H), 1.63 (s, 2H), 1.51 (s, 9H), 1.45-1.37 (m, 6H). Three unobserved exchangeable protons.

d) (R)-4-hydroxy-7-(1-hydroxy-2-(4-(2-(4-(2-isopropylthiazol-5-carbonyl) ))-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethoxy)benzylamino)ethyl)benzo[d]thiazole-2 (3H )-ketodi-trifluoroacetate

Adding trifluoroacetic acid (1.2 mL) to (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl (4-(2-(4-(2-isopropylthiazol-5-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethoxy) Benzyl) carbamic acid tert-butyl ester (Example 114, step c) (0.39 g) in DCM (4.7 mL). The resulting mixture was stirred at RT for 15 min. Toluene (30 mL) was added and the mixture was evaporated in vacuo (×3). The residue was purified by HPLC (Phenomenex , gradient: 5 to 40% acetonitrile in 0.1% aqueous trifluoroacetic acid). The product-containing fractions were combined and lyophilized to give the title compound as a white solid. The yield was 0.201 g.

m/z 668(M+H) ⁺

¹ H NMR (400 MHz, D ₄ -MeOH) δ 7.91 (s, 1H), 7.42 (d, J = 8.6 Hz, 2H), 7.06 (d, J = 8.6 Hz, 2H), 6.93 (d, J = 8.3 Hz, 1H), 6.71 (d, J = 8.3 Hz, 1H), 4.91-4.88 (m, 1H), 4.38 (t, J = 4.9 Hz, 2H), 4.24 - 4.15 (m, 2H), 3. 3.71 (m, 5H), 3.66-3.57 (m, 4H), 3.56-3.48 (m, 2H), 3.38-3.25 (m, 1H), 3.08-3.00 (m, 2H), 2.29-2.17 (m, 2H) ), 2.12-1.93 (brs, 1H), 1.88-1.73 (m, 2H), 1.38 (d, J = 6.7 Hz, 6H). Six unobserved exchangeable protons.

Example 115

(R)-4-hydroxy-7-(1-mercapto-2-(9-(4-(2-isopropylthiazol-5-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)nonylamino)ethyl)benzo[d]thiazole-2(3H)-one formate

a) (9-(9-Hydroxymercapto)-1-oxa-4,9-diazaspiro[5.5]undecethane-4-yl)(2-isopropylthiazol-5-yl) Ketone

Add a solution of 9-bromo-1-nonanol (0.446 g) in MeCN (3.9 mL) to (2-isopropylthiazol-5-yl)(1-oxa-4,9-diazaspiro) [5.5] undecane-4-yl)methanone (Example 114, step b) (0.395 g) and triethylamine (0.36 mL) in EtOAc. The resulting mixture was stirred at 60 ° C under argon for 16 h. After cooling to RT, the solvent was removed in vacuo. The residue was taken up in EtOAc EtOAc m. The residue was purified by EtOAc (EtOAc) elute The yield was 0.329 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.81 (s, 1H), 3.74 (s, 4H), 3.63 (t, J = 6.6Hz, 2H), 3.59 (s, 2H), 3.36-3.26 (m, 1H), 3.10-2.49 (m, 5H), 2.03-1.91 (m, 3H), 1.68 (s, 2H), 1.61-1.50 (m, 3H), 1.43 (d, J = 6.9 Hz, 6H), 1.36 -1.29 (m, 11H). An unobserved exchangeable proton.

b) (R)-4-hydroxy-7-(1-hydroxy-2-(9-(4-(2-isopropylthiazol-5-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)decylamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

Trifluoroacetic acid (0.055 mL) was added to (9-(9-hydroxyindenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4- at 0 °C under argon. (2-Isopropylthiazol-5-yl)methanone (Example 115, step a) (0.325 g) in EtOAc. The resulting mixture was stirred for 5 min and then Dess-Martin periodinane (0.458 g) was added. The mixture was stirred at RT for 1.5 h. A saturated solution of sodium thiosulfate (14 mL), EtOAc (EtOAc) The layers were separated and the aqueous extracted with EtOAc EtOAc. The combined organic layers were taken with EtOAc (EtOAc)EtOAc. Yellow oil was dissolved in anhydrous MeOH (15 mL) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (0.284 g), followed by acetic acid (0.062 mL) and Molecular sieves. After stirring for 5 min at RT, the reaction mixture was cooled to EtOAc (EtOAc m. The reaction mixture was filtered and the solvent was evaporated in vacuo. The residue was purified by HPLC (Phenomenex , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The product-containing fractions were combined and lyophilized to give the title compound as a white solid. The yield was 0.090 mg.

m/z 660(M+H) ⁺

^{1 H NMR (400MHz, D4-} MeOH) δ8.50 (s, 1H), 7.90 (s, 1H), 6.95 (d, J = 8.5Hz, 1H), 6.73 (d, J = 8.5Hz, 1H), 4.96-4.90 (m, 1H), 3.79-3.68 (m, 4H), 3.64-3.55 (m, 2H), 3.38-3.26 (m, 1H), 3.11-3.05 (m, 2H), 3.04-2.92 (m , 4H), 2.86-2.69 (m, 4H), 2.07-2.04 (m, 2H), 1.76-1.55 (m, 6H), 1.39-1.36 (m, 7H), 1.36-1.31 (m, 9H). Five unobserved exchangeable protons.

pEC ₅₀ and intrinsic activity of the intrinsic activity of ‧ lines expressed as the ratio of the maximum activity measured in each experiment of formoterol (formoterol).

Examples 116-182

a) (R)-3-fluoro-5-((4-(2,2,2-trifluoroethyl)-1-oxa-4,9-diazaspiro[5.5]undecane -9-yl)methyl)phenethyl (2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl)amine Tert-butyl carboxylic acid

2,2,2-trifluoro-1-(9-(3-fluoro-5-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] Carboxy-4-yl)ethanone (Example 41, step c) (0.8 g) in dichloromethane (50 mL) EtOAc (EtOAc) Treatment with iodosane (1.091 g), and the resulting mixture was stirred at 20 ° C for 40 minutes. The reaction mixture was treated with aq. EtOAc (EtOAc m. The mixture was extracted with EtOAc (EtOAc) (EtOAc)EtOAc. The residue was dissolved in methanol (3 mL) and added to (R)-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one, HCl (WO 2007027134, Example 1, step d) (0.780 g) in a solution of methanol (20 mL) and acetic acid (0.113 mL). This mixture was treated with sodium cyanoborohydride (0.186 g) and stirred at 20 ° C for 2 hr. Triethylamine (0.69 mL) was added followed by di-tert-butyl dicarbonate (0.689 mL) and stirred at 20 ° C for 2 hours. The solvent was removed under reduced pressure and the residue was partitioned between ethyl acetate and brine. The organic layer was dried over sodium sulfate, filtered and filtered and evaporated. The crude product was purified by silica gel chromatography eluting with 10% methanol in dichloromethane containing 1% concentrated aqueous ammonia. The pure fractions were evaporated to dryness to give the subtitle compound. The yield is 0.7g.

m/z 713.3(M+H) ⁺

b) (R)-3-(1-oxa-4,9-diazaspiro[5.5]undecane-9-ylmethyl)-5-fluorophenethyl (2-hydroxy-2- (4-Hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl)carbamic acid tert-butyl ester]

(R)-3-fluoro-5-((4-(2,2,2-trifluoroethyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)methyl)phenethyl (2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl)aminocarboxylic acid A solution of the third butyl ester (Examples 116-182, step a) (0.7 g) in 35% aqueous ammonia (15 mL) was placed at 20 ° C for 40 min. The reaction mixture was evaporated to one and a half of the original volume under reduced pressure. Water (10 mL) was added and the solution was passed through 10 g of C18 silica Cartridge which had been previously weighed with water. The column was rinsed with water (20 mL). The column was then flushed with methanol (50 mL) to bring out the product. The solvent was evaporated under reduced pressure and the residue wasjjjjjjjjjjj The yield was 0.580 g.

m/z 617(M+H) ⁺

c) Parallel Synthesis - Preparation of Examples 116 to 182

(R)-3-(1-oxa-4,9-diazaspiro[5.5]undecane-9-ylmethyl)-5-fluorophenethyl (2-hydroxy-2-(4) -Hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl)carbamic acid tert-butyl ester (Examples 116-182, step b) (496 mg) and A solution of triethylamine (244 mg) in NMP (2.4 mL) was partitioned in a total volume of 30 uL. A solution of the appropriate acid (0.01 mmol) in NMP (80 uL) was added to each portion, followed by a solution of HATU (4.9 mg) in NMP (30 uL). The reaction mixture was allowed to stand at room temperature for 18 hours. The additive nitrile (800 uL) was passed through a 'Tosic-65' resin (350 mg). The resin was washed with acetonitrile (800 uL), the combined wash solution was taken and passed through the 'Tosic-65' resin again. The resin was then washed with acetonitrile (3 mL). A solution of ammonia in methanol (2.5 mL of a 3.5 M solution) was passed through the resin, and the resulting washings were evaporated under a nitrogen stream. The residue was dissolved in formic acid (250 uL) and mixture was allowed to stand at room temperature for 18 hr. The additive nitrile (600 uL) was passed through a 'Tosic-65' resin (350 mg). The resin was washed with acetonitrile (800 uL), the combined wash solution was taken and passed through the 'Tosic-65' resin again. The resin was then washed with acetonitrile (3 mL). A solution of ammonia in methanol (2.5 mL of a 3.5 M solution) was passed through the resin, and the resulting washings were evaporated under a nitrogen stream. DMSO (200 uL) was added to the residue and the resulting solution was purified by &quot;Sunfire&quot; prep C18 column (19 x 50 mm) using acetonitrile to a gradient gradient elution of 0.1% aqueous TFA. The fractions containing the product were evaporated to give the title compound.

Examples 183 to 222

a) 9-bromofurfural

N-Ethyldiisopropylamine (23 mL) and DMSO (9.7 mL) were added to a solution of 9-bromo-1-decanol (10.0 g) in anhydrous DCM (230 mL). The mixture was cooled to -15 ° C and sulfur trioxide-pyridine complex (21.4 g) was added in 4 portions at 5 min intervals. After the addition was completed, the mixture was stirred for 15 min, then water (200 mL) was added and layered. The organic layer was washed with EtOAc EtOAc EtOAc EtOAc. The yield was 9.02 g.

¹ H NMR (400 MHz, CDCl ₃ ): δ 9.77 (t, J = 1.8 Hz, 1H), 3.40 (t, J = 6.8 Hz, 2H), 2.42 (td, J = 7.3, 1.8 Hz, 2H), 1.89 -1.80 (m, 2H), 1.66-1.57 (m, 2H), 1.58-1.26 (m, 8H).

b) (R)-9-bromoindolyl (2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl)amine Tert-butyl carboxylic acid

9-bromofurfural (Examples 183-222, Step a) (3.48 g), (R)-7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2 A solution of (3H)-ketohydrochloride (WO 2007027134, Example 1 part d) (4.54 g) and acetic acid (0.99 mL) in anhydrous DMF (91 mL) was stirred at RT for 30 min. The reaction mixture was cooled to 0 ° C and sodium triethyl sulfoxyborohydride (5.00 g) was added in 4 portions at intervals of 2 min. The resulting mixture was stirred at 0 °C for 5 min and then with RT for 1 h. Di-tert-butyl dicarbonate (4.12 g) was added to the reaction mixture, stirred for 1.5 h then cooled to 0 ° C. Ethyl acetate (200 mL) was added and the mixture was stirred vigorously 15 min. The layers were separated and the aqueous layer was extracted with ethyl acetate (3×150 mL). The combined organic layers were dried over sodium sulfate, filtered and evaporated in vacuo. The residue was purified by EtOAc EtOAc (EtOAc) elute The yield was 1.68 g.

m/z 532(M+H) ⁺

_{^{1 H NMR (400MHz, CDCl 3}} ): 67.02-6.90 (m, 1H); 6.78 (d, J = 8.4Hz, 1H), 4.92-4.87 (m, 1H), 3.63-3.27 (m, 5H), 3.20 -3.04 (m, 1H), 1.89-1.79 (m, 2H), 1.71-1.17 (m, 21H) + 3 unobservable exchangeable protons.

c) (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzene well [d]thiazole-7-yl)ethyl (9-(4-(2) , 2,2-trifluoroethenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)indenyl)-tert-butyl carbamate

(R)-9-bromoindolyl (2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl)amine A solution of tert-butyl formate (Examples 183 to 222, step b) (1.68 g) in MeCN (8 mL) was added dropwise to 2,2,2-trifluoro-1-(1-oxo-4, 9-diazaspiro[5.5]undecane-4-yl)ethanone trifluoroacetate (Example 12, step d) (1.18 g) and triethylamine (0.81 mL) in MeCN (8 mL) In the solution. The resulting mixture was stirred at 80 ° C for 18 h. The reaction was cooled to RT and the solvent was removed in vacuo. The brown residue was taken in EtOAc (EtOAc)EtOAc. The brown foam was purified by chromatography on EtOAc (EtOAc) elute The yield is 500 mg.

¹ H NMR (400 MHz, CDCl 3 ): δ 6.95 (s, 1H), 6.80 (dd, J = 8.3, 4.0 Hz, 1H), 4.93 (s, 1H), 3.79-3.26 (m, 10H), 3.03 -2.64 (m, 6H), 2.10 - 1.98 (m, 4H), 1.78-1.70 (m, 2H), 1.48 (s, 9H), 1.36-0.96 (m, 12H) plus 3 unobservable exchangeable Things.

d) (R)-9-(1-oxa-4,9-diazaspiro(5.5)undecane-9-yl)indolyl (2-hydroxy-2-(4-hydroxy-2-) Tert-butyl-2,3-dihydrobenzo[d]thiazole-7-yl)ethyl)carbamic acid tert-butyl ester

A solution of potassium carbonate (165 mg) in water (24 mL) was added to (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazole- 7-yl)ethyl (9-(4-(2,2,2-trifluoroethyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl A solution of the tert-butyl) carbamic acid tert-butyl ester (Examples 183-222, step c) (495 mg) in MeOH (24 mL). The resulting mixture was stirred at RT for 4 h. Methanol was removed by evaporation of the reaction mixture at 30 ° C under a stream of nitrogen. Water (20 mL) and brine (80 mL) wereEtOAc. The combined organic layers were dried over sodium sulfate, filtered and evaporated in vacuo. Standard colored solid residue was purified by silica gel chromatography, to 0 to 20% (in MeOH 2M NH ₃₎ / DCM eluting generate the subtitle compound as a yellow solid. The yield was 323 mg.

m/z 607(M+H) ⁺

^{1 H NMR (400MHz, DMSO)} : δ6.80-6.68 (m, 1H), 6.65 (d, J = 8.2Hz, 1H), 5.58 (br s, 1H), 4.69-4.56 (m, 1H), 3.44 (t, J = 3.9 Hz, 2H), 3.20-3.11 (m, 2H), 3.05-2.94 (m, 2H), 2.57 (t, J = 5.2 Hz, 2H), 2.50-2.46 (m, 2H), 2.40-2.31 (m, 2H), 2.25-2.13 (m, 4H), 1.77-1.68 (m, 2H), 1.45-1.02 (m, 25H) + 3 unobservable exchangeable protons.

e) Parallel Chemistry - Preparation of Examples 183 to 222

(R)-9-(1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)indolyl (2-hydroxy-2-(4-hydroxy-2-sideoxy) Tert-butyl-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl)carbamate (292 mg) and triethylamine (Examples 183-222, step d) (0.20 mL) The solution in NMP (1.14 mL) was dispensed in a total volume of 30 uL. A solution of the appropriate acid (0.01 mmol) in NMP (80 uL) was added to each portion, followed by a solution of HATU (4.0 mg) in NMP (30 uL). The reaction mixture was allowed to stand at room temperature overnight. The additive nitrile (800 uL) was passed through a 'Tosic-65' resin (350 mg). The resin was washed with acetonitrile (800 uL), the combined wash solution was taken and passed through the 'Tosic-65' resin again. The resin was then washed with acetonitrile (3 mL). Ammonia (3.5 M in methanol, 2.4 mL) was passed through a resin and the resulting washings were evaporated under a nitrogen stream. The residue was dissolved in formic acid (250 uL) and the mixture was placed at room temperature overnight. The additive nitrile (600 uL) was passed through a 'Tosic-65' resin (350 mg). The resin was washed with acetonitrile (800 uL), the combined wash solution was taken and passed through the 'Tosic-65' resin again. The resin was then washed with acetonitrile (3 mL). Ammonia (3.5 M in methanol, 2.4 mL) was passed through a resin and the resulting washings were evaporated under a nitrogen stream. DMSO (360 uL) was added to the residue and the resulting solution was purified by &quot;Sunfire&quot; prep C18 column (19 x 50 mm) using acetonitrile to a gradient gradient elution of 0.1% aqueous TFA. The fractions containing the product were evaporated to give the title compound.

Examples 223 to 263

a) (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl (4-(2-(4) -(2,2,2-trifluoroethenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethoxy)benzyl)carbamic acid Third butyl ester

(R)-Methanesulfonic acid 2-(4-((T-butoxycarbonyl(2-hydroxy-2-(4-hydroxy-2-yloxy)-2,3-dihydrobenzo[d]thiazole A solution of -7-yl)ethyl)amino)methyl)phenoxy)ethyl ester (Example 108, step c) (2.20 g) in MeCN (8.5 mL) was added dropwise at RT under argon. 2,2,2-Trifluoro-1-(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)ethanone trifluoroacetate (Example 12, step d (1.18 g) and a solution of triethylamine (0.81 mL) in MeCN (8.5 mL). The resulting mixture was stirred at 60 ° C under argon for 25 h. After cooling to RT, the solvent was removed in vacuo. The brown residue was taken in EtOAc EtOAc m. The yellow solid was purified by EtOAc (EtOAc) elute The yield was 729 mg.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ 6.88-6.50 (m, 6H), 4.86-4.64 (br s, 1H), 4.34-4.15 (br s, 3H), 4.11-3.87 (m, 2H), 3.70 -3.01 (m, 11H), 2.95-2.73 (br s, 2H), 2.04-1.81 (m, 2H), 1.56-1.37 (m, 11H) plus 3 unobservable exchangeables.

b) (R)-4-(2-(1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)ethoxy)benzyl (2-hydroxy-2- (4-Hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazole-7-yl)ethyl)carbamic acid tert-butyl ester

A solution of potassium carbonate (240 mg) in water (35 mL) was added to (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazole- 7-yl)ethyl (4-(2-(4-(2,2,2-trifluoroethyl)-1-oxa-4,9-diazaspiro[5.5]undecane- A solution of 9-yl)ethoxy)benzyl)carbamic acid tert-butyl ester (Examples 223-263, step a) (725 mg) in MeOH (35 mL). The resulting mixture was stirred at RT for 5 h. Methanol was removed by evaporation of the reaction mixture at 25 ° C under a stream of nitrogen. Water (20 mL) and brine (30 mL) were. The combined organic layers were dried over sodium sulfate, filtered and evaporated in vacuo. The residue was purified by silica gel chromatography, to 0 to 20% (in MeOH 2M NH ₃₎ / DCM eluting generate the subtitle compound as a white solid. The yield was 385 mg.

¹ H NMR (400 MHz, D ₆ -DMSO): δ 7.06-6.97 (m, 2H), 6.83 (d, J = 8.6 Hz, 2H), 6.78-6.62 (m, 2H), 5.64 (s, 1H) , 4.68 (br s, 1H), 4.24 (s, 2H), 3.98 (t, J = 5.6 Hz, 2H), 3.50 (t, 2H), 3.24 - 3.02 (m, 2H), 2.68-2.60 (m, 4H), 2.57 (s, 2H), 2.52-2.47 (m, 2H), 2.32 (td, J = 11.9, 3.0 Hz, 2H), 1.76 (d, J = 13.0 Hz, 2H), 1.43 (td, J = 10.8, 3.7 Hz, 2H), 1.31 (s, 3H), 1.22 (s, 6H) plus 3 unobservable exchangeables.

e) Parallel Chemistry - Preparation of Examples 223 to 263

(R)-4-(2-(1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)ethoxy)benzyl (2-hydroxy-2-(4) -Hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl)carbamic acid tert-butyl ester (Examples 223 to 263, step b) (297 mg) and A solution of triethylamine (0.20 mL) in NMP (1.24 mL) was partitioned in a total volume of 30 uL. A solution of the appropriate acid (0.01 mmol) in NMP (80 uL) was added to each portion, followed by a solution of HATU (4.0 mg) in NMP (30 uL). The reaction mixture was placed at RT overnight. The additive nitrile (800 uL) was passed through a 'Tosic-65' resin (350 mg). The resin was washed with acetonitrile (800 uL), the combined wash solution was taken and passed through the 'Tosic-65' resin again. The resin was then washed with acetonitrile (3 mL). Ammonia (3.5 M in methanol, 2.4 mL) was passed through a resin and the resulting washings were evaporated under a nitrogen stream. The residue was dissolved in formic acid (250 uL) and the mixture was placed at room temperature overnight. The additive nitrile (600 uL) was passed through a 'Tosic-65' resin (350 mg). The resin was washed with acetonitrile (800 uL), the combined wash solution was taken and passed through the 'Tosic-65' resin again. The resin was then washed with acetonitrile (3 mL). Ammonia (3.5 M in methanol, 2.4 mL) was passed through a resin and the resulting washings were evaporated under a nitrogen stream. DMSO (360 uL) was added to the residue and the resulting solution was purified by &quot;Sunfire&quot; prep C18 column (19 x 50 mm) using acetonitrile to a gradient gradient elution of 0.1% aqueous TFA. The fractions containing the product were evaporated to give the title compound.

One unobserved exchangeable H.

Example 264

(R)-4-hydroxy-7-(1-hydroxy-2-(2-(5-((4-(5-isopropylthiophen-3-carbonyl)-1-oxa-4,9-di) Azaspiro[5.5]undecane-9-yl)methyl)thiophen-3-yl)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetic acid salt

a) 2,2,2-trifluoro-1-(9-((4-(2-hydroxyethyl))thiophen-2-yl)methyl)-1-oxa-4,9-diazaspiro [5.5] undecane-4-yl) ethyl ketone

The TBAF (in THF 1M, 2.96mL) was added to 1- (9 - ((4- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethyl) thiophen-2-yl) methyl --1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)-2,2,2-trifluoroethyl ketone (Example 43, step a) (1.500 g, 2.96 mmol) in a stirred solution in THF (5 mL). After 1 h, the solution evaporated to a gum. Purification by silica gel chromatography, eluting with ethyl acetate: triethylamine 10:1 affords the subtitle compound. The yield is 0.25 g.

m/z 393(M+H) ⁺ (APCI+)

b) 2-(5-((4-(2,2,2-Trifluoroethenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl) Methyl)thiophen-3-yl)acetaldehyde

The sub-title compound was used in a similar manner to that described in the step (e) of Example 43 using 2,2,2-trifluoro-1-(9-((4-(2-hydroxyethyl)thiophen-2-yl)) Prepared by benzyl-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)ethanone (0.15 g) (Example 264, step a). The yield was 0.15 g.

m/z 391(M+H) ⁺ (APCI+)

c) (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl (2-(5-(( 4-(2,2,2-Trifluoroethenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)thiophen-3-yl) Ethyl) butyl methacrylate

(R)-7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one ‧ HCl (1.020 g) (WO 2007027134, Example 1, Step d) is added to 2-(5-((4-(2,2,2-trifluoroethyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 To a stirred solution of benzyl)methyl)thiophen-3-yl)acetaldehyde (Example 264, step b) (0.9 g) and EtOAc (EtOAc) After 2 min, sodium cyanoborohydride (0.290 g) was added. After 2 h, triethylamine (1.1 mL) was added, followed by BOC-anhydride (0.845 g). After 1 h, more BOC anhydride (0.4 g) and triethylamine (0.5 mL) were added. After 2 h, the solution was concentrated to ~5 mL then partitioned between ethyl acetate and saturated brine. The ethyl acetate layer was dried with sodium sulfate, filtered and evaporated in vacuo. Purification by silica gel chromatography, eluting with methanol:dichloromethane: 880 ammonia 10:90:1 to give a sub-title compound. The yield was 0.32 g.

m/z 701(M+H) ⁺ (APCI+)

d) (R)-2-(5-(1-oxa-4,9-diazaspiro[5.5]undecal-9-ylmethyl)thiophen-3-yl)ethyl (2- Hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzene well [d]thiazole-7-yl)ethyl)carbamic acid tert-butyl ester

Add 35% aqueous ammonia solution (5 mL) to (R)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl (2-(5-((4-(2,2,2-trifluoroethenyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)) Tert-butyl thiophen-3-yl)ethyl)carbamate (Example 264, step c) (0.2 g). After 40 min at 20 ° C, the solution was concentrated to ~1 mL and the sluice was taken to C18 (10 g) column eluted with water (20 mL) and then the product was eluted with methanol. The fractions containing the product were combined and evaporated in vacuo. The solid formed was dissolved in acetonitrile and evaporated in vacuo. The yield was 0.11 g.

m/z 605(M+H) ⁺ (APCI+)

e) (R)-4-hydroxy-7-(1-hydroxy-2-(2-(5-((4-(5-isopropylthiophen-3-carbonyl)-1-oxo-4,9) -diazaspiro[5.5]undecane-9-yl)methyl)thiophen-3-yl)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-three Fluoroacetate

Add HATU (0.075 g) to (R)-2-(5-(1-oxa-4,9-diazaspiro[5.5]undecal-9-ylmethyl)thiophen-3-yl Ethyl (2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)ethyl)carbamic acid tert-butyl ester (implementation) Example 264, step d) (0.1 g), 5-isopropylthiophene-3-carboxylic acid (0.028 g) and triethylamine (0.092 mL) After 1 h, the reaction mixture was partitioned between water and ethyl acetate. The ethyl acetate layer was washed with water (x2) and brine, dried over sodium sulfate, filtered and evaporated. The formed gum was dissolved in formic acid (2 mL). After 16 h, the solution was evaporated in vacuo. Acetonitrile was added and the mixture was evaporated (x 2) in vacuo. Toluene was added and the mixture was evaporated in vacuo. The mixture was dissolved in methanol, filtered and purified by preparative HPLC (Sunfire ^TM, Gradient: in aqueous 0.2% TFA in acetonitrile 10 to 40%) by. The fractions containing the pure product were combined and evaporated in vacuo. Acetonitrile (200 mL) was added and the solution was evaporated in vacuo to a gum. Repeat this process twice. Diethyl ether was added and the title compound was collected as a solid. The yield was 0.05 g.

m/z 657(M+H) ⁺ (multi mode +)

^{1 H NMR (400MHz, DMSO,} 90 ℃) δ11.29 (s, 1H), 7.49 (s, 1H), 7.41 (s, 1H), 7.16 (s, 1H), 6.96-6.88 (m, 2H), 6.76 (d, J = 10.5 Hz, 1H), 4.94-4.86 (m, 1H), 4.47 (s, 2H), 3.71-3.62 (m, 2H), 3.57-3.48 (m, 2H), 3.48-3.40 ( m, 2H), 3.31-2.91 (m, 11H), 2.11-1.95 (m, 2H), 1.80-1.63 (m, 2H), 1.28 (d, J = 6.6 Hz, 6H) plus 5 disappearable exchangeable protons .

Example 265

(R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(2-(pentan-3-yl)thiazole-4-carbonyl)-1-oxa-4,9 -diazaspiro[5.5]undecane-9-yl)methyl)phenethyl Amino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-(pentane) -3-yl)thiazol-4-yl)methanone

The sub-title compound was prepared using a procedure analogous to the procedure of step (b) of Example 82 using 2-(pentan-3-yl)thiazole-4-carboxylic acid (0.12 g) (Example 71, step d). The yield was 0.18 g.

¹ H NMR (400 MHz, DMSO, 90 ° C) δ 7.92 (s, 1H), 7.18 (t, J = 7.6 Hz, 1H), 7.11-7.04 (m, 3H), 4.24 (dd, J = 5.3, 3.5) Hz, 1H), 3.72-3.57 (m, 8H), 3.41 (s, 2H), 2.71 (t, J = 6.9 Hz, 2H), 2.41-2.24 (m, 4H), 1.82-1.66 (m, 4H) , 1.59-1.47 (m, 2H), 0.86 (t, J = 7.3 Hz, 6H) + 3 H which are blurred by the water peak.

b) (R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(2-(pentan-3-yl)thiazole-4-carbonyl)-1-oxa-4 ,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethyl Amino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

The title compound was used in a similar manner to that described in the step (c) of Example 82 (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] Monocarbonyl-4-yl)(2-(pentan-3-yl)thiazol-4-yl)methanone (0.18 g) (Example 265, step a). The crude aldehyde was dissolved in DCM instead of methanol. The yield was 0.09 g.

m/z 680(M+H) ⁺ (multi mode +)

¹ H NMR (400 MHz, DMSO, 90 ° C) δ 11.36 (s, 1H), 7.78 (s, 1H), 7.44 - 7.32 (m, 4H), 6.93 (d, J = 8.3 Hz, 1H), 6.77 (d, J = 8.3 Hz, 1H), 4.94 - 4.86 (m, 1H), 4.34 - 4.24 (m, 2H), 3.75 - 2.88 (m, 17H), 2.13-1.96 (m, 2H), 1.79-1.59 (m, 6H), 0.83 (t, J = 7.3 Hz, 6H) + 5 unobservable exchangeables.

Example 266

(R)-7-(2-(3-((4-(benzofuran-5-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl) )methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetic acid

a) benzofuran-5-yl(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl Ketone

The sub-title compound was prepared using a procedure analogous to that described in step (b) of Example 82 using benzofuran-5-carboxylic acid (0.1 g). The yield was 0.16 g. ¹ H NMR (400 MHz, DMSO, 90 ° C) δ 7.99 (d, J = 2.1 Hz, 1H), 7.67 (s, 1H), 7.60 (d, J = 7.7 Hz, 1H), 7.31 (dd, J = 8.6 , 1.7 Hz, 1H), 7.16 (t, J = 7.6 Hz, 1H), 7.10-7.02 (m, 3H), 6.97 (t, J = 1.2 Hz, 1H), 4.24 (t, J = 5.3 Hz, 1H) ), 3.68-3.58 (m, 4H), 3.52-3.34 (m, 6H), 2.70 (t, J = 6.9 Hz, 2H), 2.38-2.27 (m, 4H), 1.81-1.68 (m, 2H), 1.55-1.42 (m, 2H)

b) (R)-7-(2-(3-((4-(benzofuran-5-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetic acid

The title compound was used in a similar manner to that described in step (c) of Example 82 using benzofuran-5-yl(9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-di Azaspiro[5.5]undecane-4-yl)methanone (0.15 g) (Example 266, step a) was prepared. The crude aldehyde was dissolved in DCM instead of methanol. The yield was 0.07 g.

m/z 643(M+H) ⁺ (multi-mode +)

^{1 H NMR (300MHz, DMSO,} 90 ℃) δ11.38 (s, 1H), 8.02 (d, J = 2.1Hz, 1H), 7.71 (s, 1H), 7.63 (d, J = 8.5Hz, 1H) , 7.47-7.31 (m, 5H), 6.99-6.90 (m, 2H), 6.77 (d, J = 8.3 Hz, 1H), 4.95-4.86 (m, 1H), 4.33-4.25 (m, 2H), 3.72 - 2.94 (m, 16H), 2.14-1.98 (m, 2H), 1.77-1.53 (m, 2H) + 5 unobservable exchangeables.

Example 267

(R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(5-propylthiophene-3-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(5-propylthiophene) -3-yl)methanone

The sub-title compound was prepared using a procedure analogous to that described in step (b) of Example 82 using 5-propylthiophene-3-carboxylic acid (0.1 g). The yield was 0.15 g.

¹ H NMR (400 MHz, DMSO, 90 ° C) δ 7.44 (d, J = 1.3 Hz, 1H), 7.18 (t, J = 7.4 Hz, 1H), 7.11 - 7.04 (m, 3H), 6.87 (d, J = 1.3 Hz, 1H), 4.24 (t, J = 5.1 Hz, 1H), 3.67-3.59 (m, 4H), 3.51-3.47 (m, 2H), 3.40 (s, 4H), 2.77 (dd, J =14.7, 0.9 Hz, 2H), 2.71 (t, J = 7.0 Hz, 2H), 2.40-2.25 (m, 4H), 1.74-1.59 (m, 4H), 1.55-1.45 (m, 2H), 0.94 ( t, J = 7.3 Hz, 3H).

b) (R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(5-propylthiophene-3-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

The title compound was used in a similar manner to that described in the step (c) of Example 82 (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] Monocarbonyl-4-yl)(5-propylthiophen-3-yl)methanone (0.15 g) (Example 267, step a). The crude aldehyde was dissolved in DCM instead of methanol. The yield was 0.9 g.

m/z 651(M+H) ⁺ (multi mode +)

^{1 H NMR (300MHz, DMSO,} 90 ℃) δ11.38 (s, 1H), 7.51-7.32 (m, 5H), 6.96-6.87 (m, 2H), 6.77 (d, J = 8.3Hz, 1H), 4.94-4.87 (m, 1H), 4.35-4.25 (m, 2H), 3.63-2.93 (m, 16H), 2.76 (t, J = 7.3 Hz, 2H), 2.14-1.98 (m, 2H), 1.72 1.56 (m, 4H), 0.93 (t, J = 7.3 Hz, 3H) + 5 unobservable exchangeables.

Example 268

(R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(5-isopropylthiophene-2-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)methyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(5-isopropyl Thiophen-2-yl)methanone

The sub-title compound was prepared using a procedure similar to that described in step (b) of Example 82 using 5-isopropylthiophene-2-carboxylic acid (0.1 g). The yield was 0.18 g.

^{1 H NMR (400MHz, DMSO,} 90 ℃) δ 7.23-7.15 (m, 2H), 7.13-7.04 (m, 3H), 6.84 (d, J = 3.6Hz, 1H), 4.24 (t, J = 4.7Hz , 1H), 3.69-3.58 (m, 6H), 3.50 (s, 2H), 3.41 (s, 2H), 3.17 (seven peak, J = 6.8 Hz, 1H), 2.71 (t, J = 6.9 Hz, 2H) ), 2.39-2.30 (m, 4H), 1.75-1.67 (m, 2H), 1.56-1.47 (m, 2H), 1.29 (d, J = 6.7 Hz, 6H).

b) (R)-4-hydroxy-7-(1-hydroxy-2-(3-((4-(5-isopropylthiophene-2-carbonyl)-1-oxa-4,9-diaza) Hetero[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)benzene well [d]thiazole-2(3H)-one di-trifluoroacetate

The title compound was used in a similar manner to that described in the step (c) of Example 82 (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] Monocarbonyl-4-yl)(5-isopropylthiophen-2-yl)methanone (0.18 g) (Example 268, step a). The crude aldehyde was dissolved in DCM instead of methanol. The yield was 0.09 g.

m/z 651(M+H) ⁺ (multi mode +)

¹ H NMR (300 MHz, DMSO, 90 ° C) δ 11.37 (s, 1H), 7.45-7.33 (m, 4H), 7.24 (d, J = 3.1 Hz, 1H), 6.93 (d, J = 8.1 Hz, 1H), 6.85 (d, J = 3.7 Hz, 1H), 6.77 (d, J = 8.5 Hz, 1H), 4.96-4.86 (m, 1H), 4.36-4.26 (m, 2H), 3.75-3.63 (m) , 4H), 3.56-3.48 (m, 2H), 3.36-2.94 (m, 11H), 2.13-1.96 (m, 2H), 1.81-1.62 (m, 2H), 1.28 (d, J = 6.9 Hz, 6H) ) + 5 unobservable exchangeables.

Example 269

(R)-7-(2-(2,3-difluoro-4-(2-(4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)ethoxy)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-three Fluoroacetate

The title compound was used in a similar manner to that described in step (c) of Example 82 (9-(2-(2,3-difluoro-4-(2-hydroxyethyl)phenoxy)ethyl)-1-oxyl Preparation of hetero-4,9-diazaspiro[5.5]undec-4-yl)(2-isopropylthiazol-4-yl)methanone (0.22 g) (Example 77, step e). The crude aldehyde was dissolved in DCM instead of methanol. The yield was 0.13 g.

m/z 718(M+H) ⁺ (multi mode +)

^{1 H NMR (300MHz, DMSO,} 90 ℃) δ11.36 (s, 1H), 7.98 (d, J = 0.8Hz, 1H), 7.15-7.00 (m, 2H), 6.93 (d, J = 8.3Hz, 1H), 6.77 (d, J = 8.3 Hz, 1H), 4.93-4.86 (m, 1H), 4.46-4.38 (m, 2H), 3.74 - 2.94 (m, 19H), 2.15 - 1.97 (m, 2H) , 1.87-1.69 (m, 2H), 1.35 (d, J = 6.7 Hz, 6H) + 5 unobservable exchangeables.

Example 270

(R)-4-hydroxy-7-(1-hydroxy-2-(5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)methyl)-2-(trifluoromethyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoro Acetate

a) 2-(5-(bromomethyl)-2-(trifluoromethyl)phenyl)ethanol

Benzoyl peroxide (0.085 g) was added to 2-(5-methyl-2-(trifluoromethyl)phenyl)acetic acid (1.12 g) and N-bromosuccinimide (1.101 g) in chloro The suspension in benzene (18 mL) was added and the resulting mixture was heated at 100 ° C under nitrogen for 80 min then allowed to cool. The resulting mixture was diluted with ethyl acetate, washed three times with water, once with brine, then dried (MgSO _4), filtered, and concentrated in vacuo to give a yellow oil. The oil was dissolved in tetrahydrofuran (11 mL) and was taken in EtOAc EtOAc (EtOAc) The resulting mixture was stirred overnight at room temperature, then cooled in ice-water and quenched with methanol. The effervescent mixture was taken out of the cooling bath, stirred at room temperature for 70 minutes and then concentrated in vacuo. The residue was purified by flash chromatography on EtOAc (EtOAc) elute The yield was 0.919 g.

b) (9-(3-(2-Hydroxyethyl)-4-(trifluoromethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4 -yl)(2-isopropylthiazol-4-yl)methanone

Sub-title compound using 2-(5-(bromomethyl)-2-(trifluoromethyl)phenyl)ethanol (0.38 g) using a procedure similar to that described in step (b) of Example 6 (Step 270) Prepared in a solution of acetonitrile which was added dropwise to (2-isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane in 30 minutes. 4-yl)methanone trifluoroacetate (Example 22, step b). Purified by silica gel chromatography eluting with 5% triethylamine in ethyl acetate. The yield was 0.32 g.

^{1 H NMR (400MHz, DMSO,} 90 ℃) δ7.90 (s, 1H), 7.56 (d, J = 8.2Hz, 1H), 7.39 (s, 1H), 7.29 (d, J = 7.9Hz, 1H) , 4.44 (t, J = 5.3 Hz, 1H), 3.71-3.56 (m, 6H), 3.50 (s, 2H), 3.31 (seven peak, J = 6.9 Hz, 1H), 2.90 (t, J = 6.9 Hz) , 2H), 2.57-2.25 (m, 4H), 1.81-1.66 (m, 4H), 1.62-1.49 (m, 2H), 1.36 (d, J = 6.9 Hz, 6H).

c) 2-(5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)) Base-2-(trifluoromethyl)phenyl)acetaldehyde

Sub-title compound was used in a similar manner to that described in Example 16 Step c) (9-(3-(2-hydroxyethyl)-4-(trifluoromethyl)benzyl)-1-oxa-4,9 -Diazaspiro[5.5]undecyl-4-yl)(2-isopropylthiazol-4-yl)methanone (0.32 g) (Example 270, step b). The mixture was stirred for 90 min after the addition of Dess-Martin periodinane. The yield was 0.38 g.

d) (R)-4-hydroxy-7-(1-hydroxy-2-(5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diaza) Heterospiro[5.5]undecane-9-yl)methyl)-2-(trifluoromethyl)phenethylamino)ethyl)benzo[d]thiazole-2(3H)-one di- Trifluoroacetate

The title compound was used in a similar manner to that described in Example 18, step g), using 2-(5-((4-(2- isopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] Preparation of solution of undecane-9-yl)methyl)-2-(trifluoromethyl)phenyl)acetaldehyde (0.3 g) (Example 270, step c) in methanol (3 mL) . After the addition of sodium cyanoborohydride, the mixture was stirred at room temperature for 3.75 hours. The solution was concentrated to a volume ~3 mL, diluted with THF and washed with a mixture of brine and saturated aqueous sodium hydrogen carbonate. The organic phase was dried over anhydrous magnesium sulfate and evaporated The residue was co-evaporated twice from acetonitrile then EtOAc (EtOAc) (EtOAc) The fractions containing the product were combined, concentrated in vacuo and co-evaporated three times from acetonitrile to give a colourless residue. The residue was triturated with diethyl ether to give a solid, which was taken, filtered, washed with diethyl ether and dried in vacuo. The yield was 0.115 g.

m/z 720(M+H) ⁺ (APCI+)

^{1 H NMR (400MHz, DMSO,} 90 ℃) δ7.93 (s, 1H), 7.77 (d, J = 8.2Hz, 1H), 7.61-7.53 (m, 2H), 6.94 (d, J = 8.2Hz, 1H), 6.77 (d, J = 8.5 Hz, 1H), 4.91 (dd, J = 8.6, 4.7 Hz, 1H), 4.27-4.08 (m, 2H), 3.76-3.59 (m, 6H), 3.48-2.85 (m, 11H), 2.03-1.88 (m, 2H), 1.78-1.63 (m, 2H), 1.35 (d, J = 6.9 Hz, 6H). Six unobserved exchangeable protons.

Example 271

(R)-7-(2-(3-((2,2-dimethyl-4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoroacetate

a) 1-Benzyl-4-((2-methyla11y1amino)methyl)piperidin-4-ol

6-Benzyl-1-oxa-6-azaspiro[2.5]octane (2g) and 2-methylprop-2-en-1-amine HCl (2g) and Hunig's base (3.44mL) The mixture in ethanol (30 mL) was heated at 70 ° C for 18 hours. The mixture was cooled to room temperature and the solvent was evaporated under reduced pressure. The residue was partitioned between EtOAc EtOAc (EtOAc)EtOAc. The yield was 2.70 g.

m/z 275(M+H) ⁺ (APCI+)

b) 9-benzyl-2,2-dimethyl-1-oxa-4,9-diazaspiro[5.5]undecane-4-carboxylic acid tert-butyl ester

Concentrated sulfuric acid (12 ml) was added to 1-benzyl-4-((2-methylallylamino)methyl)piperidin-4-ol (Example 271, step a) (2.2 g). Leave at room temperature for 2 hours. The reaction mixture was treated with ice/water (100 mL) and then portionwise with sodium bicarbonate until the mixture was basic. Acetonitrile (50 mL) was added followed by BOC-anhydride (1.925 g) and the mixture was stirred at room temperature for 18 hours. The reaction mixture was extracted with EtOAc (EtOAc)EtOAc. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.8 g.

m/z 375(M+H) ⁺ (APCI+)

c) 2,2-dimethyl-1-oxa-4,9-diazaspiro[5.5]undecane-4-carboxylic acid tert-butyl ester

9-benzyl-2,2-dimethyl-1-oxa-4,9-diazaspiro[5.5]undecane-4-carboxylic acid tert-butyl ester dissolved in ethanol (50 mL) Example 271, a mixture of step b) (0.8 g) and 10% palladium on carbon (0.5 g) was treated with ammonium formate (0.8 g) and the reaction mixture was heated under reflux for 30 min. The mixture was cooled to room temperature and filtered through celite. The solvent was evaporated and the residue was taken to aq. The yield was 0.55 g.

^{1 H NMR (400MHz, DMSO,} 90 ℃) δ3.25 (s, 2H), 3.19 (s, 2H), 2.84-2.76 (m, 2H), 2.59-2.50 (m, 2H), 1.51-1.42 (m , 4H), 1.41 (s, 9H), 1.13 (s, 6H). 1 unobserved exchangeable substance.

d) 9-(3-(2-Hydroxyethyl)benzyl)-2,2-dimethyl-1-oxa-4,9-diazaspiro[5.5]undecane-4-carboxylic acid Third butyl ester

2-(3-(Bromomethyl)phenyl)ethanol (0.416 g) was added in portions over 1 hour to 2,2-dimethyl-1-oxa-4,9-diazaspiro[5.5] A solution of the first alkyl monocarbane-4-carboxylate (Example 271, step c) (0.55 g) and triethylamine (0.809 mL) in acetonitrile (20 mL). The mixture was stirred at 20 ° C for 2 hours. The solvent was evaporated under reduced pressure. EtOAc m. The crude product was purified by flash chromatography eluting with EtOAc (EtOAc) elute The yield was 0.78 g.

¹ H NMR (400 MHz, DMSO, 90 ° C) δ 7.21 - 7.04 (m, 4H), 4.24 (t, J = 5.5 Hz, 1H), 3.66-3.59 (m, 2H), 3.43 (s, 2H), 3.25 (s, 2H), 3.19 (s, 2H), 3.00 (s, 2H), 2.71 (t, J = 7.0 Hz, 2H), 2.32 - 2.24 (m, 2H), 1.64-1.50 (m, 4H) , 1.40 (s, 9H), 1.13 (s, 6H).

e) 2-(3-((2,2-Dimethyl-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl)phenyl)ethanol

Adding trifluoroacetic acid (5 mL) to 9-(3-(2-hydroxyethyl)benzyl)-2,2-dimethyl-1-oxa-4,9-diazaspiro[5.5] A solution of the mono- hexane- carboxylic acid tert-butyl ester (Example 271, step d) (0.78 g) in DCM (20 mL), and the reaction mixture was allowed to stand at 20 ° C for 30 minutes. Toluene (40 mL) was added, and the solvent was evaporated evaporated. The residue was dissolved in water (30 mL) and washed with ethyl acetate. The aqueous mixture was then extracted with EtOAc (EtOAc)EtOAc. The yield is 0.4g.

m/z 319(M+H) ⁺ (APCI+)

f (9-(3-(2-hydroxyethyl)benzyl)-2,2-dimethyl-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl (2-methylthiazol-4-yl)methanone

Sub-title compound using 2-(3-((2,2-dimethyl-1-oxa-4,9-diazaspiro[5.5] eleven) using the method described in the step (a) of Example 22 Preparation of carnamid-9-yl)methyl)phenyl)ethanol (0.2 g) (Example 271, step e) and 2-methylthiazole-4-carboxylic acid (0.09 g). The reaction mixture was stirred for 18 h and the solvent used for chromatography was 2% methanol in dichloromethane containing 1% triethylamine. The yield was 0.16 g.

m/z 444.1(M+H) ⁺ (APCI+)

g) (R)-7-(2-(3-((2,2-dimethyl-4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaza) Spiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one di-trifluoro Acetate

The title compound was used in a similar manner to that described in the step (d) of Example 22 (9-(3-(2-hydroxyethyl)benzyl)-2,2-dimethyl-1-oxa-4,9- Preparation of diazaspiro[5.5]undecyl-4-yl)(2-methylthiazol-4-yl)methanone (0.15 g) (Example 271, step f). The mixture was stirred for 3 h after the addition of sodium cyanoborohydride, and 2-methyltetrahydrofuran was added instead of THF. The yield was 0.12 g.

m/z 652(M+H) ⁺ (multi-mode +)

^{1 H NMR (400MHz, DMSO,} 90 ℃) δ11.27 (s, 1H), 7.93 (s, 1H), 7.43-7.32 (m, 4H), 6.93 (d, J = 8.5Hz, 1H), 6.77 ( d, J = 8.2 Hz, 1H), 4.94-4.89 (m, 1H), 4.29 (s, 2H), 3.68 (s, 2H), 3.62 (s, 2H), 3.28-3.10 (m, 8H), 3.06 - 2.97 (m, 2H), 2.67 (s, 3H), 1.95-1.73 (m, 4H), 1.19 (s, 6H). 5 unobserved exchangeables.

Example 272

(R)-5-(2-(3-((4-(5-ethylthiophen-3-carbonyl)-2,2-dimethyl-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetate

a) (5-ethylthiophen-3-yl)(9-(3-(2-hydroxyethyl)benzyl)-2,2-dimethyl-1-oxa-4,9-diaza Spiro[5.5]undecane-4-yl)methanone

Sub-title compound using 2-(3-((2,2-dimethyl-1-oxa-4,9-diazaspiro[5.5] eleven) using the procedure described in step (f) of Example 271. Carboxane-9-yl)methyl)phenyl)ethanol (0.2 g) (Example 271, step e) and 5-ethylthiophene-3-carboxylic acid. The yield was 0.22 g.

m/z 457.1(M+H) ⁺ (APCI+)

b) (R)-5-(2-(3-((4-(5-ethylthiophen-3-carbonyl)-2,2-dimethyl-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-8-hydroxyquinoline-2(1H)-one di-trifluoroacetate

The title compound was used in a similar manner to that described in Example 23 (5-ethylthiophen-3-yl)(9-(3-(2-hydroxyethyl)benzyl)-2,2-dimethyl-1- Prepared by oxa-4,9-diazaspiro[5.5]undec-4-yl)methanone (0.22 g) (Example 272, step a). The mixture was stirred for 3 h after the addition of sodium triethoxysulfonate in step a). The yield was 0.085 g.

m/z 659(M+H) ⁺ (multi mode +)

¹ H NMR (400 MHz, DMSO, 90 ° C) δ 8.16 (d, J = 10.0 Hz, 1H), 7.48 (d, J = 1.5 Hz, 1H), 7.44 - 7.29 (m, 4H), 7.13 (d, J = 8.2 Hz, 1H), 6.99 (d, J = 8.2 Hz, 1H), 6.91 (d, J = 1.0 Hz, 1H), 6.55 (d, J = 10.0 Hz, 1H), 5.37-5.32 (m, 1H), 4.24 (s, 2H), 3.52-3.40 (m, 4H), 3.28 (t, J = 8.3 Hz, 2H), 3.20-2.99 (m, 8H), 2.81 (q, J = 7.4 Hz, 2H) ), 1.93-1.65 (m, 4H), 1.25 (t, J = 7.4 Hz, 3H), 1.18 (s, 6H). 6 unobserved exchangeables.

Example 273

(R)-8-hydroxy-5-(1-hydroxy-2-(5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro) (5.5) undecane-9-yl)methyl)-2-(trifluoromethyl)phenethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

(R) -5- (2- amino-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) -8-hydroxy-quinolin -2 (1H) - one (0.15g) (WO 2004106333) A solution of methanol (2 mL) was taken in acetic acid (0.021 mL) and stirred for 10 min. Subsequent addition of 2-(5-((4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)) a solution of 2-(trifluoromethyl)phenyl)acetaldehyde (Example 270, step c) (0.190 g) in MeOH (3 mL). Cooled in ice-water and treated with sodium cyanoborohydride (0.038 g). The cooling bath was removed and the mixture was stirred at room temperature for 2.75 hours. The solution was concentrated to a volume of ~3 mL, diluted with EtOAc (20 mL) and washed with brine (10 mL) and sat. The organic phase was dried over anhydrous magnesium sulfate and evaporated The residue was co-evaporated twice from EtOAc (EtOAc) (EtOAc) The mixture was concentrated in vacuo and EtOAc EtOAc (EtOAc)EtOAc The product containing fractions were concentrated in vacuo and co-evaporated three times from acetonitrile to give a colourless residue. The residue was triturated with EtOAc (EtOAc)EtOAc. The yield was 0.133 g.

m/z 714(M+H) ⁺ (APCI+)

¹ H NMR (400 MHz, DMSO, 90 ° C) δ 8.17 (d, J = 10.0 Hz, 1H), 7.93 (s, 1H), 7.77 (d, J = 8.2 Hz, 1H), 7.62-7.53 (m, 2H) ), 7.14 (d, J = 8.2 Hz, 1H), 7.00 (d, J = 8.2 Hz, 1H), 6.55 (d, J = 10.0 Hz, 1H), 5.34 (dd, J = 9.1, 4.0 Hz, 1H) ), 4.29-4.10 (m, 2H), 3.76-3.59 (m, 6H), 3.49-2.87 (m, 11H), 2.05-1.87 (m, 2H), 1.80-1.62 (m, 2H), 1.34 (d) , J = 6.9 Hz, 6H). Six unobserved exchangeable protons.

Example 274

(R)-8-hydroxy-5-(1-hydroxy-2-(3-((4-(2-(pentan-3-yl)thiazole-4-carbonyl)-1-oxa-4,9 -diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

a) 2-(3-((4-(2-(pentan-3-yl)thiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 9-yl)methyl)phenyl)acetaldehyde

Sub-title compound was used in a similar manner to that described in Example 16 Step c) using (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5] Monocarbonyl-4-yl)(2-(pentan-3-yl)thiazol-4-yl)methanone (0.16 g) (Example 265, step a). The yield was 0.24 g.

m/z 470(M+H) ⁺ (APCI+)

b) (R)-8-Hydroxy-5-(1-hydroxy-2-(3-((4-(2-(pentan-3-yl)thiazole-4-carbonyl)-1-oxa-4 ,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetate

The title compound was used in a similar manner to that described in Example 273 using 2-(3-(4-(2-(pentan-3-yl)thiazole-4-carbonyl)-1-oxa-4,9-diaza Preparation of heterospiro[5.5]undecane-9-yl)methyl)phenyl)acetaldehyde (Example 274, step a). The yield was 0.097 g.

m/z 674(M+H) ⁺ (APCI+)

¹ H NMR (400 MHz, DMSO, 90 ° C) δ 8.15 (d, J = 10.0 Hz, 1H), 7.95 (s, 1H), 7.47-7.28 (m, 4H), 7.13 (d, J = 8.2 Hz, 1H), 6.99 (d, J = 7.9 Hz, 1H), 6.55 (d, J = 9.7 Hz, 1H), 5.39-5.28 (m, 1H), 4.33-4.11 (m, 2H), 3.79-3.56 (m , 6H), 3.50-2.84 (m, 11H), 2.11-1.90 (m, 2H), 1.83-1.58 (m, 6H), 0.84 (t, J = 7.3 Hz, 6H). Six unobserved exchangeable protons.

Example 275

(R)-8-hydroxy-5-(1-mercapto-2-(1-(3-((4-(5-isopropylthiophen-3-carbonyl)-1-oxa)-4,9- Diazaspiro[5.5]undecane-9-yl)methyl)phenyl)-2-methylpropan-2-ylamino)ethyl)quinoline-2(1H)-one di-three Fluoroacetate

a) 3-(2-( First three -butoxycarbonylamino)-2-methylpropyl)benzoic acid

The sub-title compound was prepared using a procedure analogous to the procedure of step b) of Example 53 using methyl 3-(2-(tris-butoxycarbonylamino)-2-methylpropyl)benzoate. The reaction mixture was stirred at 20 ° C for 18 hours and then at 40 ° C for 10 hours. The mixture is partitioned between water and diethyl ether and does not require final wet milling.

m/z 292(MH) ⁺ (APCI+)

b) tert-butyl 1-(3-(hydroxymethyl)phenyl)-2-methylpropan-2-ylcarbamate

The subtitle compound of Example 16 using similar step a) of the method using 3- (2- (tertiary - butoxycarbonyl) -2-methylpropyl) benzoic acid (2.68 g of) was prepared. During the addition, the reaction mixture was cooled in an ice bath and then stirred at 20 ° C for 2 hours. The yield is 2.5g.

¹ H NMR (400 MHz, DMSO) δ 7.23 - 7.13 (m, 2H), 7.06 (s, 1H), 6.98 (d, J = 7.4 Hz, 1H), 6.23 (s, 1H), 5.09 (t, J = 5.6 Hz, 1H), 4.45 (d, J = 5.6 Hz, 2H), 2.89 (s, 2H), 1.42 (s, 9H), 1.14 (s, 6H).

c) (3-(2-Amino-2-methylpropyl)phenyl)methanol

Trifluoroacetic acid (5 mL) was added to 1-butyl 3-(3-(hydroxymethyl)phenyl)-2-methylpropan-2-ylcarbamate (Example 275, step b) (0.7 g The resulting solution was allowed to stand at 20 ° C for 30 minutes in a solution of DCM (20 mL). Toluene (40 mL) was added and the solvent was evaporated under reduced pressure. The residue was partitioned between DCM and 1M aqueous sodium hydroxide. The combined organics were dried over sodium sulfate, filtered and evaporated <RTI ID=0.0> The yield was 0.440 g.

m/z 180(M+H) ⁺ (APCI+)

^{1 H NMR (400MHz, DMSO)} δ7.23 (t, J = 8.0Hz, 1H), 7.19-7.12 (m, 2H), 7.04 (d, J = 8.6Hz, 1H), 4.47 (s, 2H), 2.58 (s, 2H), 0.99 (s, 6H). 3 unobserved exchangeables.

d) (R)-8-(benzyloxy)-5-(1-( third -butyldimethylmethylalkyloxy)-2-(1-(3-(hydroxymethyl)phenyl)-2-methylpropan-2-ylamino)ethyl)quinoline-2 (1H )-use

(R) -8- (benzyloxy) -5- (2-bromo-l- (- butyldimethylsilyl silicon alkyloxy) ethyl) quinolin -2 (1H) - one (0.518 g) and (3-(2-Amino-2-methylpropyl)phenyl)methanol (0.19g) (Example 275, Step c) and sodium iodide (0.159g) and Hunig's base (0.555mL) The mixture in acetonitrile (3 mL) was refluxed under nitrogen for 2 days. Additional (3-(2-Amino-2-methylpropyl)phenyl)methanol (0.19 g) and Hunig's base (0.185 mL) were added and heating was continued for one day under reflux. The reaction mixture was cooled to rt. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.44 g.

m/z 587(M+H) ⁺ (APCI+)

e) (R)-5-(1-(Third-butyldimethylmethylalkyloxy)-2-(1-(3-(hydroxymethyl)phenyl)-2-methylpropane-2 -ylamino)ethyl)-8-hydroxyquinoline-2(1H)-one

(R) -8- (benzyloxy) -5- (l- (tertiary - butyl dimethyl silicone alkyloxy) -2- (l- (3- (hydroxymethyl) phenyl) - 2-methylpropan-2-ylamino)ethyl)quinolin-2(1H)-one (Example 275, step d) (0.44 g) and palladium on carbon (10%) (80 mg) in ethanol ( The mixture in 30 mL) was vigorously stirred under 5 bar of hydrogen pressure for 3 hours. The catalyst was filtered off and the solvent was evaporated under reduced pressure to give subtitle compound. The yield was 0.37 g.

m/z 497(M+H) ⁺ (APCI+)

f (R)-third-butyl carbonate 5-(1-(tris-butyldimethylmethylalkyloxy)-2-(1-(3-(hydroxymethyl)phenyl)-2- Methylpropan-2-ylamino)ethyl)-2-yloxy-1,2-dihydroquinolin-8-yl ester

(R)-5-(1-( Third -butyldimethylmethylalkyloxy)-2-(1-(3-(hydroxymethyl)phenyl)-2-methylpropan-2-yl Amino)ethyl)-8-hydroxyquinoline-2(1H)-one (Example 275, step e) (0.37 g) and BOC-anhydride (0.163 g) and triethylamine (0.208 mL) in ethanol ( The solution in 20 mL) was allowed to stand at room temperature for 18 hours. The solvent was removed under reduced pressure to give the subtitle compound. The yield was 0.44 g.

m/z 597(M+H) ⁺ (APCI+)

g) (R)-tris-butyl carbonate 5-(1-(t-butyldimethylmethylalkyloxy)-2-(1-(3-methylindenyl)-2-yl Propane-2-ylamino)ethyl)-2-yloxy-1,2-dihydroquinolin-8-yl ester

Adding manganese dioxide (0.6 g) to (R)-tri-butyl carbonate 5-(1-(Third-butyldimethylsilyloxy)-2-(1-(3-(hydroxyl) Methyl)phenyl)-2-methylpropan-2-ylamino)ethyl)-2-yloxy-1,2-dihydroquinolin-8-yl ester (Example 275, step f) (0.44 g) in a solution of dichloromethane (30 mL), and the mixture was stirred vigorously at 20 ° C for 3 hours. The reaction mixture was filtered through EtOAc (EtOAc)EtOAc. The yield was 0.42 g.

m/z 595(M+H) ⁺ (APCI+)

h)(R)-3-(2-(2-( third -butyldimethylmethylalkyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino)-2-methylpropyl Benzaldehyde

(R)-tris-butyl carbonate 5-(1-(Third-butyldimethylsilyloxy)-2-(1-(3-methylindenyl)-2-methylpropane a solution of 2-ylamino)ethyl)-2-oxoyl-1,2-dihydroquinolin-8-yl ester (Example 275, step g) (0.42 g) in methanol (15 mL) Treated with ammonia (35% in water) (2 mL). The reaction mixture was allowed to stand at 20 ° C for 3 hours. The solvent was removed under a stream of nitrogen and the residue was partitioned between ethyl acetate and brine. The aqueous layer was extracted with EtOAc (EtOAc)EtOAc. The yield was 0.34 g.

m/z 495(M+H) ⁺ (APCI+)

i) (5-isopropylthiophen-3-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone

The subtitle compound was prepared using a procedure similar to that described in steps 22 and b) of Example 22 using 5-isopropylthiophene-3-carboxylic acid. In step b), the residue is not purified by chromatography. The crude product was azeotroped (x 2) with acetonitrile. The residue was partitioned between EtOAc EtOAc EtOAc. The yield is 0.4g.

m/z 309(M+H) ⁺ (APCI+)

j) (R)-5-(1-(Third-butyldimethylmethylalkyloxy)-2-(1-(3-((4-(5-isopropylthiophen-3-carbonyl))) -1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenyl)-2-methylpropan-2-ylamino)ethyl)-8 -hydroxyquinolin-2(1H)-one

The title compound was prepared using a similar embodiment the sub-step in Example 14 b) using the method of (R) -3- (2- (2- ( tertiary - butyl dimethyl silicone alkyloxy) -2- (8-hydroxy - 2-Phenoxy-1,2-dihydroquinolin-5-yl)ethylamino)-2-methylpropyl)benzaldehyde (0.32 g) (Example 275, step h) and (5- Isopropylthiophen-3-yl)(1-oxa-4,9-diazaspiro[5.5]undec-4-yl)methanone (0.21 g) (Example 275, step i)) preparation. The crude product was purified by flash chromatography on EtOAc (EtOAc) elute The pure fractions were evaporated to dryness to give the subtitle compound. The yield was 0.31 g.

m/z 787(M+H) ⁺ (APCI+)

k) (R)-8-hydroxy-5-(1-hydroxy-2-(1-(3-((4-(5-isopropylthiophen-3-carbonyl)-1-oxo-4,9) -diazaspiro[5.5]undecane-9-yl)methyl)phenyl)-2-methylpropan-2-ylamino)ethyl)quinoline-2(1H)-one di- Trifluoroacetate

The title compound was prepared using similar to Example 23, step b) using the method of (R) -5- (1- (third - butyldimethylsilyl silicon alkyloxy) -2- (1- (3 - ((4 -(5-isopropylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenyl)-2-methyl Propane-2-ylamino)ethyl)-8-hydroxyquinolin-2(1H)-one (0.31 g) (Example 275, step j). The yield was 0.18 g.

m/z 673(M+H) ⁺ (multi-mode +)

¹ H NMR (400 MHz, DMSO, 90 ° C) δ 9.98 (s, 1H), 8.14 (d, J = 10.0 Hz, 1H), 7.47 (s, 1H), 7.44 - 7.40 (m, 2H), 7.36- 7.30 (m, 2H), 7.20 (d, J = 8.2 Hz, 1H), 7.01 (d, J = 8.2 Hz, 1H), 6.89 (s, 1H), 6.58 (d, J = 9.7 Hz, 1H), 5.37-5.31 (m, 1H), 4.26 (s, 2H), 3.68-3.63 (m, 2H), 3.55-3.51 (m, 2H), 3.21-2.98 (m, 11H), 2.01 (s, 2H), 1.69 (s, 2H), 1.31-1.24 (m, 12H) plus 5 unobservable exchangeables.

Example 276

8-hydroxy-5-((R)-1-hydroxy-2-((R)-1-(3-((4-(5-isopropylthiophen-3-carbonyl)-1-oxa-4) ,9-diazaspiro[5.5]undecane-9-yl)methyl)phenyl)propan-2-ylamino)ethyl)quinoline-2(1H)-one di-trifluoroacetic acid salt

a) 1-(3-((Third-butyldimethylmethylalkyloxy)methyl)phenyl)propan-2-one

(3-Bromo-benzyloxy) (tertiary-butyl) - dimethyl Silane (16.72g), isopropenyl acetate (9.2 mL), tri-n-butyltin methoxide (24mL), palladium acetate (II) (0.625g And a mixture of tris(o-tolyl)phosphine (1.70 g) in dry toluene (65 mL) was heated at reflux <RTI ID=0.0> The mixture was diluted with ethyl acetate (130 mL) and aq. The diatomaceous earth was added, and the suspension was filtered through celite, and the residue was washed thoroughly with ethyl acetate. The combined filtrate and washings were dried over anhydrous magnesium sulfate, EtOAc (EtOAc m. Dissolved to give a yellow oil of the sub-title compound. The yield was 13.71 g.

¹ H NMR (400MHz, CDCl ₃ ) δ 7.30 (t, J = 7.6 Hz, 1H), 7.23 (d, J = 7.7 Hz, 1H), 7.16 (s, 1H), 7.09 (d, J = 7.4 Hz) , 1H), 4.73 (s, 2H), 3.68 (s, 2H), 2.14 (s, 3H), 0.94 (s, 9H), 0.10 (s, 6H).

b (3-((R)-2-((R)-1-phenylethyl)amino)propyl)phenyl)methanol hydrochloride

1-(3-(( Tertiary -butyldimethylsilyloxy)methyl)phenyl)propan-2-one (Example 276, Step a) (6.85 g) in dichloromethane (240 mL) The solution was treated with acetic acid (1.27 mL), (R)-(+)-1-phenylethylamine (2.83 mL) and sodium triethyloxyborohydride (7.09 g). Stir overnight at room temperature. The mixture was quenched by the addition of saturated aqueous sodium bicarbonate (120 mL) over 5 min. The biphasic mixture was stirred vigorously for 1.75 hours and then separated. The aqueous phase was extracted with more dichloromethane, and the combined organic phase was dried (MgSO _4), filtered, and concentrated to give an oil. The oil was purified by flash chromatography on EtOAc (EtOAc) elute elute elute elute The oil was dissolved in HCl / MeOH (1M, 20 mL). The resulting gel was wet milled with methanol/diethyl ether to give a crystal slurry which was stirred for 30 minutes. The solid was removed by filtration, washed with diethyl ether and crystallised from methanol to diethyl ether to yield white crystals. The yield was 1.9 g.

m/z 270(M+H) ⁺ (APCI+)

¹ H NMR (400 MHz, DMSO) δ 9.84 (br s, 1H), 9.21. (br s, 1H), 7.69 (d, J = 6.9 Hz, 2H), 7.53-7.40 (m, 3H), 7.23 (t) , J = 7.6 Hz, 1H), 7.16 (d, J = 7.7 Hz, 1H), 6.98 (s, 1H), 6.88 (d, J = 7.4 Hz, 1H), 5.16 (t, J = 5.6 Hz, 1H) ), 4.69-4.55 (m, 1H), 4.43 (d, J = 5.4 Hz, 2H), 3.42-3.30 (m, 1H), 3.04-2.91 (m, 1H), 2.61-2.47 (m, 1H), 1.63 (d, J = 6.7 Hz, 3H), 1.10 (d, J = 6.7 Hz, 3H).

c) (R)-(3-(2-Aminopropyl)phenyl)methanol

(3-((R)-2-((R)-1-Phenylethyl)amino)propyl)phenyl)methanol hydrochloride (Example 276, step b) (1.92 g), 20% carbon A mixture of palladium hydroxide (0.757 g) and ammonium formate (2.28 g) in ethanol (30 mL) was stirred at 75 ° C for 1.5 hr then cooled. The suspension was filtered through celite and the residue was washed thoroughly with ethanol. The combined filtrate and wash are concentrated in vacuo to yield a pale yellow gum of the desired product and mixture of dehydroxy analogs. The yield was 1.54 g. Use directly.

d) (R)-1-(3-(hydroxymethyl)phenyl)propan-2-ylaminocarboxylic acid tert-butyl ester

A solution of di-tert-butyl dicarbonate (2.034 g) in THF (5 mL) was added dropwise to impure (R)-(3-(2-aminopropyl)phenyl)methanol over 20 min. 276, step c) (1.54 g) in THF (10 mL) and water (10 mL). The resulting mixture was stirred at room temperature over the weekend before triethylamine (3.90 mL) was added. More di-tert-butyl dicarbonate (2.013 g) in THF was added dropwise over 30 minutes. The resulting mixture was stirred at room temperature overnight then concentrated in vacuo to remove THF. Formation of an oily aqueous phase was extracted twice with ethyl acetate, and the combined extracts were dried (MgSO _4), filtered and concentrated onto flash silicon dioxide in vacuo. The residue was purified by flash chromatography eluting EtOAc EtOAc The yield was 0.9 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ7.29 (t, J = 7.6Hz, 1H), 7.25-7.17 (m, 2H), 7.11 (d, J = 7.4Hz, 1H), 4.68 (s, 2H) , 4.37 (br s, 1H), 3.91 (br s, 1H), 2.85 (dd, J = 12.7, 5.0 Hz, 1H), 2.66 (dd, J = 13.3, 7.4 Hz, 1H), 1.42 (s, 9H) ), 1.09 (d, J = 6.7 Hz, 3H). An unobserved exchangeable proton.

e) 8-(Benzyloxy)-5-((R)-1-(tris-butyldimethylmethylalkyloxy)-2-((R)-1-(3-(hydroxyl) Phenyl)propan-2-ylamino)ethyl)quinoline-2(1H)-one

(R)-1-butyl 3-(3-(hydroxymethyl)phenyl)propan-2-ylcarbamate (Example 276, step d) (0.78 g) in dichloromethane (20 mL) The solution was treated with trifluoroacetic acid (5 mL) and stirred at room temperature for 40 min. Toluene was added and the solution was concentrated in vacuo. The residue was partitioned between 1 M aqueous NaOH and DCM and the phases separated. The aqueous phase was extracted twice with DCM multiple, after bonding the organic extracts were dried (MgSO _4), and concentrated in vacuo to give (R) - as a colorless oil (3- (2-aminopropyl) phenyl) methanol, It slowly crystallized as a white solid (0.442 g) upon standing. (R)-(3-(2-Aminopropyl)phenyl)methanol (0.223 g), (R)-8-(benzyloxy)-5-(2-bromo-1-( third- a mixture of butyldimethylsilyloxy)ethyl)quinolin-2(1H)-one (0.72g), sodium iodide (0.219g) and Hunig's base (0.77mL) in acetonitrile (3mL) Heat overnight under reflux under nitrogen. More (R)-(3-(2-Aminopropyl)phenyl)methanol (0.219 g), Hunig's base (0.51 mL) and acetonitrile (2 mL) were added and the mixture was heated under reflux for the next night. The cooled reaction mixture was diluted with ethyl acetate, washed twice with water, once with brine, then dried (MgSO ₄₎ and concentrated in vacuo to silicon dioxide in the flash. The resulting powder was purified by flash chromatography on EtOAc (EtOAc) elute The yield was 0.525 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ9.37 (br s, 1H), 8.46 (d, J = 9.7Hz, 1H), 7.69-7.56 (m, 5H), 7.39-7.22 (m, 4H), 7.17 (d, J = 8.2 Hz, 1H), 7.12 (d, J = 6.7 Hz, 1H), 6.82 (d, J = 10.0 Hz, 1H), 5.37 (d, J = 2.1 Hz, 2H), 5.26 (dd , J = 7.4, 5.6 Hz, 1H), 4.82 (s, 2H), 3.24 (dd, J = 11.4, 7.6 Hz, 1H), 3.05 (q, J = 6.5 Hz, 1H), 2.93 (dd, J = 11.4, 5.5 Hz, 1H), 2.84 (dd, J = 13.6, 7.0 Hz, 1H), 2.74 (dd, J = 13.2, 6.8 Hz, 1H), 1.25 (d, J = 6.2 Hz, 3H), 1.04 ( s, 9H), 0.24 (s, 3H), 0.00 (s, 3H). Two unobserved exchangeable protons.

f) 5-((R)-1-(Third-butyldimethylmethylalkyloxy)-2-((R)-1-(3-(hydroxymethyl)phenyl)propane-2- Amino)ethyl)-8-hydroxyquinoline-2(1H)-one

8- (benzyloxy) -5 - ((R) -1- ( III - alkyloxy silicon butyldimethylsilyl) -2 - ((R) -1- (3- ( hydroxymethyl) Phenyl)propan-2-ylamino)ethyl)quinolin-2(1H)-one (Example 276, step e) (0.52 g) and 10% carbon on palladium catalyst (0.102 g) in ethanol ( The mixture in 36 mL) was hydrogenated at room temperature and under a hydrogen pressure of 5 bar for 1.5 hours, then hydrogenated overnight at room temperature under a hydrogen pressure of 1 bar. The mixture was filtered through celite, the catalyst was washed well with ethanol, and the combined filtrate and washings were concentrated in vacuo to give sub-title compound. The yield was 0.42 g. The substance is used directly in the next steps.

m/z 483(M+H) ⁺ (APCI+)

g) (R)-2-(8-(Third-butoxycarbonyloxy)-2-oxooxy-1,2-dihydroquinolin-5-yl)-2-(tri-butyl) Dimethyl decyloxy)ethyl ((R)-1-(3-(hydroxymethyl)phenyl)propan-2-yl)carbamic acid tert-butyl ester

5-((R)-1-( Third -butyldimethylmethylalkyloxy)-2-((R)-1-(3-(hydroxymethyl)phenyl)propan-2-ylamine Ethyl)-8-hydroxyquinolin-2(1H)-one (Example 276, step f) (0.42 g) was dissolved in ethanol (25 mL) with triethylamine (0.26 mL) The third butyl ester (0.201 g) was treated and stirred at room temperature for 1.5 hours. More di-tert-butyl dicarbonate (0.203 g) and triethylamine (0.13 mL) were added and the mixture was stirred at room temperature overnight then concentrated in vacuo to give subtitle compound. The yield was 0.609 g. The substance is used directly in the next steps.

m/z 683(M+H) ⁺ (APCI+)

h) (R)-2-(8-(Third-butoxycarbonyloxy)-2-oxooxy-1,2-dihydroquinolin-5-yl)-2-(tri-butyl) Dimethyl decyloxy)ethyl ((R)-1-(3-methylnonylphenyl)propan-2-yl)carbamic acid tert-butyl ester

(R)-2-(8-(Third-butoxycarbonyloxy)-2-oxooxy-1,2-dihydroquinolin-5-yl)-2-(tris-butyl dimethyl Tert-butyloxyalkyl)ethyl ((R)-1-(3-(hydroxymethyl)phenyl)propan-2-yl)carbamic acid tert-butyl ester (Example 276, step g) (0.609 g Dissolved in dichloromethane (30 mL), treated with manganese (IV) oxide (0.791 g), and the resulting suspension was stirred at room temperature over the weekend. More manganese (IV) oxide (0.801 g) was added and the suspension was stirred for another 4 hours. The mixture was filtered through celite, and the residue was washed with DCM. The yield was 0.521 g. The substance is used directly in the next steps.

m/z 681(M+H) ⁺ (APCI+)

i (R)-2-(Third-butyldimethylmethylalkyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl ((R)-1-(3-methylnonylphenyl)propan-2-yl)carbamic acid tert-butyl ester

(R)-2-(8-(Third-butoxycarbonyloxy)-2-oxooxy-1,2-dihydroquinolin-5-yl)-2-(tris-butyl dimethyl Tertiary alkyl (oxy)oxy)ethyl ((R)-1-(3-methylnonylphenyl)propan-2-yl)carbamic acid tert-butyl ester (Example 276, step h) (0.521 g) suspended Treatment with methanol (20 mL) in EtOAc (2 mL) (EtOAc) The mixture was concentrated by blowing a stream of nitrogen, and the residue was partitioned between ethyl acetate, water and brine, and the phases were separated. The aqueous phase was extracted with ethyl acetate, the combined organic phase was washed with brine, dried (MgSO _4), and concentrated in vacuo to give the sub-title compound. The yield was 0.476 g. The substance is used directly in the next steps.

m/z 581(M+H) ⁺ (APCI+)

j) (R)-2-(Third-butyldimethylsilyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)B (R)-1-(3-((4-(5-isopropylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)methyl)phenyl)propan-2-yl)carbamic acid tert-butyl ester

(R)-2-(Third-butyldimethylsilyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl ( (R)-1-butyl 3-(3-methylnonylphenyl)propan-2-yl)carbamate (Example 276, step i) (0.476 g) as (5-isopropylthiophene-3) -yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)methanone (Example 275, step i) (0.296 g), N-methyl-2 Treatment with pyrrolidone (20 mL), acetic acid (0.052 mL) and sodium triethyloxyborohydride (0.294 g), The solution was poured into a mixture of saturated sodium bicarbonate solution and water and extracted with diethyl ether three times. The combined extracts were washed with water and brine, dried (MgSO _4), concentrated in vacuo, then purified by flash chromatography on silicon dioxide by to _{1: 5: 94 NEt 3:} MeOH: DCM eluting produce subtitle Compound. The yield was 0.471 g.

m/z 874(M+H) ⁺ (APCI+)

k) 8-hydroxy-5-((R)-1-hydroxy-2-((R)-1-(3-((4-(5-isopropylthiophen-3-carbonyl)-1-oxa) -4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenyl)propan-2-ylamino)ethyl)quinoline-2(1H)-one di-three Fluoroacetate

(R)-2-(Third-butyldimethylsilyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethyl ( (R)-1-(3-((4-(5-isopropylthiophene-3-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl) a solution of tert-butyl methyl)phenyl)propan-2-yl)carbamate (Example 276, step j) (0.465 g) in formic acid (25 mL). Dilute and concentrate in vacuo. The residue was dissolved in a mixture of water and EtOAc (1 mL), filtered, and purified by preparative HPLC (Sunfire, gradient: 5 to 50% acetonitrile in 0.2% aqueous TFA). The product containing fractions were concentrated in vacuo and co-evaporated three times from acetonitrile to give a colourless residue. The residue was triturated with EtOAc (EtOAc)EtOAc. The yield was 0.234 g.

m/z 659(M+H) ⁺ (APCI+)

¹ H NMR (400 MHz, DMSO, 90 ° C) δ 8.20-8.13 (m, 1H), 7.48 (d, J = 1.0 Hz, 1H), 7.46-7.31 (m, 4H), 7.20-7.13 (m, 1H) ), 7.00 (d, J = 8.2 Hz, 1H), 6.90 (s, 1H), 6.56 (d, J = 9.7 Hz, 1H), 5.37 (t, J = 6.4 Hz, 1H), 4.29 (s, 2H) ), 3.70-3.62 (m, 2H), 3.62-3.50 (m, 3H), 3.49-3.36 (m, 2H), 3.30-2.98 (m, 8H), 2.81-2.71 (m, 1H), 2.12-1.93 (m, 2H), 1.81-1.60 (m, 2H), 1.28 (d, J = 6.7 Hz, 6H), 1.20-1.13 (m, 3H). Six unobserved exchangeable protons.

Example 277

(R)-N-cyclohexyl-N-(2-(2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazol-7-yl)) Amino)ethyl)-3-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 9-yl)methyl)phenylethoxy)propane decylamine di-trifluoroacetate

a) 3-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl Phenyl ethoxy) propionic acid tert-butyl ester

Add Triton-B (0.094 mL) to (9-(3-(2-hydroxyethyl)benzyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4- (2,4-Methylthiazol-4-yl)methanone (Example 6, step b) (1.72 g) in EtOAc (30 mL). The mixture was concentrated in vacuo and cooled to 0 °C. Tributyl acrylate (0.671 mL) was added over 1 minute, and the mixture was stirred at room temperature for 20 hours. The reaction was diluted with EtOAc (EtOAc)EtOAc. The substance is used directly in the next steps.

b) 3-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl Phenylethoxy)propionic acid

3-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)benzene Ethyl ethoxy) tert-butyl propionate (Example 277, step a) was dissolved in DCM (10 mL) and TFA (2mL). The resulting mixture was stirred for 2 h then evaporated. The residue was azeotroped with toluene and redissolved in MeCN (10 mL). This material was applied to an SCX column (50 g, Varian) that had been pre-wetted with MeCN. The column was washed with MeCN (100 mL) and dissolved in MeCN (1:4, 100 mL). The eluent is evaporated to give the sub-title compound. The substance is used directly in the next steps.

c) N-cyclohexyl-N-(2,2-dimethoxyethyl)-3-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4, 9-diazaspiro[5.5]undecane-9-yl)methyl)phenylethoxy)propanylamine

3-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)benzene Ethoxy)propionic acid (Example 277, step b) was dissolved in DMF (10 mL) and N-(2,2-dimethoxyethyl)cyclohexaneamine (WO 2008075025) (0.93 g), Triethylamine (2.88 mL) and T3P (3.95 mL). The reaction was stirred overnight and partitioned between water (100 mL) andEtOAc. The layers were separated and the aqueous was extracted with ethyl acetate (2×100 mL). The combined organics were washed with EtOAc EtOAc m. The residue was purified by flash chromatography on EtOAc EtOAc (EtOAc): . The yield was 0.9 g.

¹ H NMR (400 MHz, DMSO, 90 ° C) δ 7.85 (s, 1H), 7.26-7.05 (m, 4H), 4.46-4.37 (m, 1H), 3.68-3.55 (m, 11H), 3.43 (s) , 2H), 3.33 - 3.22 (m, 8H), 2.77 (t, J = 6.9 Hz, 2H), 2.68 (s, 3H), 2.56 (t, J = 6.7 Hz, 2H), 2.38-2.32 (m, 4H), 1.78-1.65 (m, 4H), 1.63-1.42 (m, 6H), 1.33-1.19 (m, 2H), 1.15-1.00 (m, 2H).

d) N-cyclohexyl-3-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9 -yl)methyl)phenethyloxy)-N-(2-o-oxyethyl)propane decylamine

Add p-toluenesulfonic acid (1384 mg) to N-cyclohexyl-N-(2,2-dimethoxyethyl)-3-(3-((4-(2-methylthiazole-4-carbonyl)) -1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl)phenethyloxy)propanoguanamine (Example 277, step c) (683 mg) in DCM (10 mL) in solution, and the resulting mixture was stirred at ambient temperature for 4 h. Sodium bicarbonate solution (saturated, 10 mL) was added cautiously and the mixture was stirred until bubbling ceased (10 min). The reaction was diluted with DCM (50 mL) and aqueous layer was separated. The organics were washed with aq. EtOAc (EtOAc)

m/z 611(M+H) ⁺ (APCI+)

e) (R)-N-cyclohexyl-N-(2-(2-hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydrobenzo[d]thiazole-7-yl) Ethylamino)ethyl)-3-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undeccarbon) Alkan-9-yl)methyl)phenylethoxy)propane decylamine di-trifluoroacetate

N-Cyclohexyl-3-(3-((4-(2-methylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9- Add a solution of (meth) phenylethoxy)-N-(2-oxoethyl)propanoguanamine (Example 277, step d) (0.122 g) in methanol (3 mL) to (R) -7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one, HCl (WO 2007027134, Example 1, step d) (0.079 g) and In a mixture of acetic acid (0.011 mL). The resulting mixture was stirred for 5 min and then cooled to 0 °C. Sodium cyanoborohydride (0.019 g) was then added and the mixture was warmed to ambient temperature and stirred for 2 h. The solvent was evaporated and the residue was purified by flash column chromatography eluting with EtOAc (EtOAc: EtOAc) The fractions containing the product were combined, evaporated and purified by HPLC (Sunfire, gradient: 5 to 40% acetonitrile in 0.2% aqueous TFA). The title compound was obtained as a white solid. The yield was 0.085 g.

m/z 821(M+H) ⁺ (multi mode +)

¹ H NMR (400 MHz, DMSO, 90 ° C) δ 11.26 (s, 1H), 7.90 (s, 1H), 7.41-7.28 (m, 4H), 6.93 (d, J = 8.2 Hz, 1H), 6.77 ( d, J = 8.2 Hz, 1H), 4.89 (dd, J = 8.7, 4.1 Hz, 1H), 4.29 (s, 2H), 3.74 - 3.43 (m, 13H), 3.29 - 2.98 (m, 8H), 2.83 (t, J = 6.7 Hz, 2H), 2.71-2.56 (m, 5H), 1.98-1.01 (m, 14H). 5 unobserved exchangeables.

Example 278

(R)-4-hydroxy-7-(1-hydroxy-2-(2-(4-((4-(2-isopropylthiazol-4-carbonyl)-1-oxo-4,9-di) Azaspiro[5.5]undecane-9-yl)methyl)thiophen-2-yl)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-trifluoroacetic acid salt

a) third -butyl (2-(5-chlorothien-2-yl)ethoxy)dimethyl decane

N -chlorosuccinimide (0.83 g) was added in portions to tris-butyldimethyl(2-(thiophen-2-yl)ethoxy)decane (1.5 g) (Step 4 in Example 4) The solution in chloroform (50 mL) was heated to reflux for 3 days. The yellow solution was diluted with EtOAc (EtOAc)EtOAc. The residue was purified by silica gel chromatography eluting with iso -hexane. The fractions containing the product are combined and evaporated in vacuo to give the sub-title compound as a clear oil. The yield is 1g.

_{^{1 H NMR (300MHz, CDCl 3}} ) δ6.69 (d, J = 3.5Hz, 1H), 6.57-6.54 (m, 1H), 3.75 (t, J = 6.5Hz, 2H), 2.89 (t, J = 6.4 Hz, 2H), 0.87 (s, 9H), 0.01 (s, 6H)

b) 5-(2-( third -butyldimethylmethylalkyloxy)ethyl)-2-chlorothiophene-3-carbaldehyde

Tri -butyl(2-(5-chlorothiophen-2-yl)ethoxy)dimethyloxane (0.8 g) (Example 278, step a) was added dropwise to butyllithium in 5 min. 2.5 M, 7 mL of alkane and 2,2,6,6-tetramethylpiperidine (0.73 mL) in THF (25 mL). The resulting mixture was stirred for 2 hr and DMF (0.67 mL) was added. The mixture was stirred for another 1 hr and allowed to warm to RT. The reaction was cautiously poured into HCl solution (0.5M, EtOAc) The combined organic solution was washed with EtOAc (EtOAc)EtOAc. The oil formed was purified by chromatography on silica gel eluting with EtOAc (EtOAc) EtOAc. The fractions containing the product are combined and evaporated in vacuo to give the sub-title compound as a clear oil. The yield was 0.8 g.

_{^{1 H NMR (400MHz, CDCl 3}} ) δ9.98 (s, 1H), 7.06-7.04 (m, 1H), 3.79 (t, J = 6.0Hz, 2H), 2.95-2.85 (m, 2H), 0.90 ( s, 9H), 0.05 (s, 6H)

c) 5-(2-(Third-butyldimethylmethylalkyloxy)ethyl)thiophene-3-carbaldehyde

5-(2-( Third -butyldimethylsilyloxy)ethyl)-2-chlorothiophene-3-carbaldehyde (0.8 g) (Example 278, step b) in ethanol (50 mL) A mixture of ethylamine (0.91 mL) was stirred vigorously with 10% carbon on palladium catalyst (0.28 g) under a hydrogen pressure of 4 bar for 18 hours. The mixture was filtered through celite and the pad was washed with ethanol (50mL). The combined filtrate and washings were evaporated and azeotroped with toluene (20 mL) to yield a yellow oil. The oil was dissolved in DCM (100 mL) and EtOAc (EtOAc) The mixture was filtered through celite (br.) and filtered (EtOAc) The combined filtrate and washings were evaporated in vacuo to give a sub-title compound as a yellow oil. The yield was 0.6 g.

_{^{1 H NMR (300MHz, CDCl 3}} ) δ9.80 (s, 1H), 7.93-7.90 (m, 1H), 3.80 (t, J = 6.2Hz, 2H), 2.99 (t, J = 5.9Hz, 2H) , 0.88 (s, 9H), 0.01 (s, 6H) + a fuzzy H due to solvent peaks

d) (9-((5-(2-(Third-butyldimethylsilyloxy)ethyl)thiophen-3-yl)methyl)-1-oxa-4,9-diaza Heterospiro[5.5]undecane-4-yl)(2-isopropylthiazol-4-yl)methanone

5-(2-( Third -butyldimethylsilyloxy)ethyl)thiophene-3-carbaldehyde (0.6 g) (Example 278, step c) was added to N-methyl-2-pyrrole (2-Isopropylthiazol-4-yl)(1-oxa-4,9-diazaspiro[5.5]undecane-4 in a mixture of ketone (5 mL) and acetic acid (0.13 mL) Methyl ketone (0.94 g) (Example 22, step b) and stirred for 30 min. Sodium triethoxy borohydride (0.71 g) was then added and the mixture was stirred overnight. The reaction was poured into water (100 mL), EtOAc (EtOAc) The combined organic solution was washed with water (3×100 mL The residue was purified by EtOAc (EtOAc):EtOAc:EtOAc:EtOAc The fractions containing the product are combined and evaporated in vacuo to give the sub-title compound as a clear oil. The yield was 0.8 g.

564(M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D ₆ -DM So, 90 ° C) δ 7.91 (s, 1H), 6.97 (s, 1H), 6.77 (s, 1H), 3.79 (t , J = 6.3 Hz, 2H), 3.69-3.55 (m, 4H), 3.41-3.27 (m, 3H), 2.92 (td, J = 6.3, 0.8 Hz, 2H), 2.42-2.23 (m, 6H), 1.75-1.64 (m, 2H), 1.59-1.47 (m, 2H), 1.37 (d, J = 6.9 Hz, 6H), 0.89 (s, 9H), 0 (s, 6H)

e) (9-((5-(2-Hydroxyethyl)thiophen-3-yl)methyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl (2-isopropylthiazol-4-yl)methanone

Tetrabutylammonium fluoride (in tetrahydrofuran 1M, 2.1mL) was added to (9 - ((5- (2- (Third - butyldimethylsilyl silicon alkyloxy) ethyl) thiophen-3-yl Methyl)-1-oxa-4,9-diazaspiro[5.5]undecethane-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 278, step d) (0.80 g) in a solution in tetrahydrofuran (10 mL). After 2 h, the solution was evaporated EtOAc m m m m m m m The layers were separated and the aqueous extracted with ethyl acetate (2×50 mL). The combined organic solution was washed with brine (50 mL) dried over sodium sulfate. The residue was purified by silica gel chromatography eluting with 20:1 ethyl acetate: triethylamine. The fractions containing the product are combined and evaporated in vacuo to give a sub-title compound. The yield was 0.61 g.

450 (M+H) ⁺ (APCI) ¹ H NMR (400 MHz, D _{6 -} DMSO, 90 ° C) δ 7.90 (s, 1H), 6.94 (s, 1H), 6.74 (s, 1H), 4.41 (t) , J = 5.1 Hz, 1H), 3.72-3.55 (m, 4H), 3.41-3.24 (m, 3H), 3.00 (s, 2H), 2.88 (t, J = 6.7 Hz, 2H), 2.43 - 2.26 ( m, 6H), 1.75-1.30 (m, 10H)

f) (R)-4-hydroxy-7-(1-hydroxy-2-(2-(4-(4-(2-isopropyl)thiazol-4-carbonyl)-1-oxa-4,9 -diazaspiro[5.5]undecane-9-yl)methyl)thiophen-2-yl)ethylamino)ethyl)benzo[d]thiazole-2(3H)-one di-three Fluoroacetate

(9-((5-(2-Hydroxyethyl)thiophen-3-yl)methyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)( 2-isopropylthiazol-4-yl)methanone

(0.35 g) (Example 278, step e) EtOAc m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m m Stir at RT for 1 hour. The reaction mixture was treated with aq. sodium sulphate (10 mL) and EtOAc (EtOAc) The mixture was extracted with ethyl acetate (2×30 mL). The combined organic solution was washed with EtOAc EtOAc (EtOAc m. The residue was dissolved in methanol (5 mL) followed by ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.15 g) and the mixture was stirred for 5 min before cooling in an ice bath. Sodium cyanoborohydride (0.07 g) was then added and the mixture was warmed to RT and stirred overnight. The solvent was evaporated in vacuo. Purification by gel chromatography, eluting with a gradient of 94.5:5:0.5 to 89:10:1 DCM:methanol: '880' aqueous ammonia. The fractions containing the product were combined and evaporated in vacuo. It was further purified by preparative HPLC (Sunfire ^TM, Gradient: aqueous 0.2% TFA in 10 to 30% acetonitrile). The title compound was obtained as a white solid. The yield was 0.10 g.

m/z 658(M+H) ⁺ (multi mode)

¹ H NMR (500 MHz, D ₆ -DMSO) δ 11.68 (s, 1H), 10.42-10.01 (m, 2H), 9.06 (s, 1H), 8.86 (s, 1H), 8.04 (s, 1H), 7.59 (s, 1H), 7.03 (s, 1H), 6.92 (d, J = 8.4 Hz, 1H), 6.79 (d, J = 8.3 Hz, 1H), 6.52 (s, 1H), 4.94 - 4.86 (m) , 1H), 4.42-4.23 (m, 2H), 3.82-3.48 (m, 6H), 3.34 - 2.91 (m, 11H), 2.14 - 2.02 (m, 2H), 1.83-1.59 (m, 2H), 1.33 (d, J = 6.8 Hz, 6H)

Example 279

(R)-4-hydroxy-7-(1-hydroxy-2-(9-(4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)-3,3-dimethylhydrazinoamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

a) 8-(Third-butyl dimethyl decyloxy)-2,2-dimethyloctanoate ethyl ester

n-Butyllithium (32.8 mL) was added to a solution of diisopropylamine (1M in toluene, 11.64 mL) in THF (60 mL) over 15 min. The reaction mixture was cooled to -78 ° C, then ethyl isobutyrate (10 mL) was added dropwise over 15 min. The mixture was stirred at -78 ℃ 1 hour before dropwise added (6-bromo-hexyloxy) - third - butyldimethyl Silane (23 mL), and the reaction mixture was stirred at -78 deg.] C for another 60 minutes. The cooling bath was removed and the mixture was stirred with EtOAc EtOAc EtOAc m. The combined organic layers were washed with water (250 mL) dried over magnesium sulfate. Purification by gel chromatography eluting with 0 to 40% ethyl acetate in cyclohexane afforded a sub-title compound as a light amber liquid. The yield is 20g.

¹ H NMR (400 MHz, CDCl ₃ ): δ 4.11 (q, J = 7.1 Hz, 2H); 3.59 (t, J = 6.6 Hz, 2H); 1.54-1.45 (m, 4H); 1.35-1.18 (m) , 9H); 1.15 (s, 6H); 0.89 (s, 9H); 0.05-0.03 (m, 6H).

b) 8-(( third -butyldimethylmethylalkyloxy)-2,2-dimethyloctane-1-ol

Ethyl 8-(t-butyldimethylmethyl decyloxy)-2,2-dimethyloctanoate (Example 279, step a) (20 g) in dry diethyl ether (300 mL) It was cooled to 0 ° C, then diisobutylaluminum hydride (1 M in toluene, 133 mL) was added dropwise. The reaction mixture was stirred at 0 &lt;0&gt;C for 1 h then quenched with saturated aqueous sodium succinate (EtOAc &lt The mixture was extracted with EtOAc (EtOAc (EtOAc)EtOAc. Purification by gel chromatography eluting with 0 to 20% ethyl acetate in cyclohexane afforded the subtitle compound. The yield was 10.11 g.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ3.60 (t, J = 6.6Hz, 2H); 3.31 (s, 2H); 1.56-1.46 (m, 2H); 1.38-1.18 (m, 8H); 0.89 (s, 9H); 0.86 (s, 6H); 0.06-0.03 (m, 6H) plus an unobservable exchangeable.

c) (E)-T-butyl(9-methoxy-7,7-dimethylindole-8-alkenyloxy)dimethylnonane and (Z)-tertiary-butyl (9) a mixture of -methoxy-7,7-dimethylindole-8-alkenyloxy)dimethyl decane

Toluene (5.9 mL) was added to a solution of dimethyl hydrazine (4.98 mL) in anhydrous dichloromethane (60 mL). Add 8-(Third-butyldimethylsilyloxy)-2,2-dimethyloctane-1-ol dissolved in dichloromethane (20 mL) dropwise (Example 279, step b) (10.11 g), and the mixture was stirred for 1 hour. Triethylamine (19.4 mL) was then added and stirred for 1 hour. The cooling bath was removed and the mixture was allowed to warm to rt then DCM (15 mL). 1 N HCl was added dropwise to dissolve the salt present in the reaction mixture and the layers were separated. The aqueous layer was extracted with EtOAc EtOAc (EtOAc)EtOAc. The residue was filtered over silicon dioxide short plunger, using diethyl ether as eluent to give 8- (tertiary - butyl - dimethyl - Si alkyloxy) -2,2-dimethyl - oct aldehydes yellow Liquid, which was used without further purification.

Potassium tert-butoxide (15.3 g) and chlorinated (methoxymethyl)triphenylphosphonium (48 g) were cooled to 0 ° C in anhydrous THF (350 mL) and stirred for 30 min. 8-( Third -butyl-dimethyl-decyloxy)-2,2-dimethyl-octanal (10.03 g) was dissolved in anhydrous THF (150 mL) and added to the reaction mixture, which Stir at 0 ° C for 1 hour. The mixture was allowed to warm to room temperature and stirred for 30 min then a saturated sodium bicarbonate solution (300 mL). The combined organic layers were washed with EtOAc (EtOAc m.

Cyclohexane (300 mL) was added and the mixture was stirred for 5 min. The solvent was decanted and the process was repeated. Evaporation of the cyclohexane layer produced a yellow liquid with some white solid. The solid was filtered off, then the filtrate was purified by chromatography eluting with EtOAc EtOAc Further purification was achieved by dissolving the material (3.2 g) in acetone (50 mL), sodium iodide (3.05 g), followed by Merrifield resin (6.4 g) and stirring the mixture over the weekend. The resin was filtered and washed with acetone (100 mL), DCM (50mL) and methanol (50mL). The filtrate was evaporated and taken up in EtOAc EtOAc EtOAc (EtOAc) a mixture. The yield was 2.6 g.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ6.16 (d, J = 12.9Hz, 0.6H); 5.69 (d, J = 7.0Hz, 0.4H); 4.73 (d, J = 12.9Hz, 0.6H) ;4.09 (d, J=7.0 Hz, 0.4H); 3.61-3.56 (m, 2H); 3.53-3.47 (m, 3H); 1.56-1.45 (m, 2H); 1.38-1.16 (m, 8H); 1.05 (d, J = 10.1 Hz, 3H); 0.97 (s, 3H); 0.92 - 0.86 (m, 9H); 0.06 - 0.02 (m, 6H).

d) (E)-9-methoxy-7,7-dimethylindole-8-en-1-ol and (Z)-9-methoxy-7,7-dimethylindole-8- Mixture of alken-1-ol

(E) - tertiary - butyl (9-methoxy-7,7-dimethyl-enoic yloxy) dimethyl Silane (Example 279, step c) (2.57g) was dissolved Tetrahydrofuran (60 mL), tetrabutylammonium fluoride (16.4 mL) was added, and the mixture was stirred at room temperature for 4 hr. The reaction mixture was quenched with water (50 mL) andEtOAcEtOAc The organic layer was dried with EtOAc EtOAc EtOAc m. The yield was 1.2g.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ6.16 (d, J = 12.9Hz, 0.5H); 5.70 (d, J = 7.0Hz, 0.5H); 4.74 (d, J = 12.9Hz, 0.5H) ; 4.11 (d, J = 7.0 Hz, 0.5H); 3.68-3.61 (m, 2H); 3.52 (s, 1.5H); 3.49 (s, 1.5H); 1.62-1.51 (m, 2H); 1.40- 1.21 (m, 8H); 1.07 (s, 3H); 0.98 (s, 3H) plus an unobservable exchange.

e) 7,7-dimethyl-9-oxodecyl methanesulfonate

(E)-9-methoxy-7,7-dimethylindole-8-en-1-ol and (Z)-9-methoxy-7,7-dimethylindole-8-ene- A mixture of 1-alcohols (Example 279, step d) (1.6 g) was dissolved in dichloromethane (15 mL) and triethylamine (1.33 mL) was added and the mixture was cooled to 0 ° C under argon. Methanesulfonium chloride (0.68 mL) was added dropwise over 5 minutes, and the reaction mixture was stirred at 0 ° C for 15 min and then at room temperature over 1 hr. The reaction mixture was washed with EtOAc (EtOAc m. Purification by chromatography on silica gel eluting with 0 to 100% ethyl acetate in hexanes to give the subtitle compound as a colourless oil. The yield was 1.2g.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ9.85-9.81 (m, 1H); 4.25-4.18 (m, 2H); 3.00 (s, 3H); 2.25 (m, 2H); 1.80-1.69 (m, 2H); 1.47-1.37 (m, 2H), 1.31 (s, 6H); 1.05-1.01 (m, 6H).

f) 9-(4-(2-Isopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)-3,3- Dimethylfurfural

(2-Isopropyl-thiazol-4-yl)-(1-oxa-4,9-diaza-spiro[5.5]undec-4-yl)-methanone (0.262 g) (as in the example) Step 102 c was prepared from trifluoroacetate) dissolved in acetonitrile (4 mL) and triethylamine (0.22 mL). This solution was then added to a solution of 7,7-dimethyl-9-oxiranyl methanesulfonate (Example 279, step e) (0.291 g) in acetonitrile (4 mL). °C heating overnight. The solvent was removed under vacuum. Purification by silica gel chromatography, eluting with 0 to 10% methanol in DCM to give the subtitle compound as a colourless oil. The yield was 0.361 g.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ9.80-9.77 (m, 1H); 7.81 (s, 1H); 4.12-3.80 (m, 2H); 3.73 (s, 3H); 3.68-3.58 (m, 1H); 3.34-3.22 (m, 1H); 2.90-2.33 (m, 6H); 2.21 (d, J = 3.17 Hz, 2H); 1.99-1.65 (m, 6H); 1.63-1.47 (m, 2H) ; 1.37 (d, J = 6.9 Hz, 6H); 1.31-1.20 (m, 6H); 1.00 (s, 6H).

g) (R)-4-hydroxy-7-(1-hydroxy-2-(9-(4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)-3,3-dimethyldecylamino)ethyl)benzo[d]thiazole-2(3H)-one, formate

Acetic acid (0.035 mL) was added to 9-(4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl -3,3-dimethylfurfural (Example 279, step f) (195 mg) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d] Thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (161 mg) in a stirred solution of methanol (10 mL), and one minute, sodium cyanoborohydride (51 mg) The reaction mixture was stirred at room temperature for 1.5 h. The mixture was concentrated and then partitioned between THF (12 mL) and EtOAc (EtOAc:EtOAc) The layers were separated and the organic phase dried over sodium sulfate filtered and evaporated. The formed material was purified by preparative HPLC (Phenomenex , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The fractions containing the product were combined and lyophilized to give the title compound as a white solid. The yield was 0.038 g.

m/z 688(M+H) ⁺

¹ H NMR (400 MHz, D ₆ -DMSO, 80 ° C): δ 8.12 (s, 1.6H); 7.89 (s, 1H); 6.88 (d, J = 8.3 Hz, 1H); 6.70 (d, J = 8.3 Hz, 1H); 4.59 (t, J = 6.2 Hz, 1H); 3.64 (d, J = 9.5 Hz, 6H); 2.75 (d, J = 6.7 Hz, 2H); 2.61-2.53 (m, 2H) ; 2.40-2.21 (m, 6H); 1.72-1.63 (m, 2H); 1.58-1.47 (m, 2H); 1.43-1.30 (m, 10H); 1.29-1.09 (m, 8H); 0.82 (s, 6H) Add a signal that is obscured by the solvent and four unobserved exchangeables.

Example 280

(R)-4-mercapto-7-(1-hydroxy-2-(10-(4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)nonan-2-ylamino)ethyl)benzo[d]thiazole-2(3H)-one formate

a) 10-bromodecane-2-ol

To a solution of 9-bromo-nonanol (1.7 g) in DCM (100 mL) was added diisopropylethylamine (3.91 mL) followed by DMSO (1.62 mL) and the mixture was cooled to -14 ° C then added portionwise Sulfur trioxide pyridine complex (3.64 g). The mixture was stirred for 30 minutes and then allowed to stand overnight. Water was added to separate the phases. The organic layer was washed with 1M aqueous sodium hydrogen sulfate (×3), saturated aqueous sodium hydrogen carbonate and brine. The organic phase was separated and dried using a hydrophobic frit, followed by evaporation to yield a yellow liquid (2.32 g).

Methyllithium (1.6 M in diethyl ether, 2.1 mL) was added dropwise at EtOAc. After 45 minutes, the mixture was poured onto 2M sulfuric acid with stirring and the phases were separated. The organic layer was washed with aq. Purification by gel chromatography eluting with 0 to 30% ethyl acetate in cyclohexane afforded the title compound. The yield was 0.286 g.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ3.83-3.75 (m, 1H); 3.41 (t, J = 6.9Hz, 2H); 1.90-1.80 (m, 2H); 1.50-1.37 (m, 6H) ;1.36-1.26 (s, 6H); 1.19 (d, J = 6.2 Hz, 3H) plus an unobserved exchangeable substance.

b) (9-(9-Hydroxymercapto)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-isopropylthiazol-4-yl) Ketone

(2-Isopropyl-thiazol-4-yl)-(1-oxa-4,9-diaza-spiro[5.5]undec-4-yl)-methanone trifluoroacetate (Examples 22, step b) (500 mg) was applied to a methanol-wet SCX-2 column, washed with methanol, then dissolved with 2N ammonia in methanol and evaporated to give a free base (276 mg). This was mixed with 10-bromodecane-2-ol (Example 280, Step a, 280 mg) and triethylamine (0.247 mL) in acetonitrile (10 mL). The solvent was evaporated, the crude was purified eluting elut elut elut The yield was 0.298 g.

¹ H NMR (400 MHz, D ₆ -DMSO): δ 8.99 (s, 1H); 8.00 (s, 1H); 4.23 (d, J = 4.7 Hz, 1H); 3.78-3.44 (m, 8H); -2.77 (m, 4H); 2.08-1.92 (m, 2H); 1.76-1.48 (m, 4H); 1.30 (d, J = 6.9 Hz, 7H); 1.26-1.16 (m, 12H); 0.98 (d) , J = 6.1 Hz, 3H).

c) 10-(4-(2-Isopropylthiazole-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)decane-2- ketone

(9-(9-Hydroxymercapto)-1-oxa-4,9-diazaspiro[5.5]undecethane-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 280, step b) (287 mg) EtOAc m. After the mixture was stirred for 5 minutes, Dess-Martin periodinane (392 mg) was added. The reaction mixture was stirred at room temperature for 5 hours, then an additional amount of &quot;Dess-Martin periodinane (392mg) Saturated sodium thiosulfate solution (20 mL), a saturated aqueous solution of sodium hydrogencarbonate (20 mL) and ethyl acetate were added, and the mixture was stirred, and then the phases were separated. The aqueous layer was extracted with EtOAc (×2) and the combined organics were washed with saturated aqueous sodium hydrogen carbonate. Acetic acid (0.053 mL) was added. The yield was 0.338 g.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ7.85 (s, 1H); 4.07-3.72 (m, 5H); 3.36-3.21 (m, 2H); 2.96-2.82 (m, 3H); 2.47-2.37 ( m, 4H); 2.12 (s, 3H); 2.11-2.02 (m, 2H); 1.85-1.75 (m, 2H); 1.62-1.49 (m, 3H); 1.46-1.36 (m, 7H); 1.20 (m, 9H).

d) (R)-4-hydroxy-7-(1-hydroxy-2-(10-(4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)decane-2-ylamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

Acetic acid (0.053 mL) was added to 10-(4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro[5.5] eleven at room temperature under nitrogen. Cycloalkan-9-yl)nonan-2-one (Example 280, Step c) (335 mg) and ( R )-7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzo[ d] thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (243 mg) in anhydrous methanol with 3 Mix in a mixture of molecular sieves for 5 minutes. The mixture was cooled to 0 ° C and sodium triacetoxyborohydride (131 mg) was added. The reaction mixture was stirred and warmed to room temperature overnight. The mixture was filtered and evaporated. Purified by preparative HPLC (Phenomenex , Gradient: 10 to 40% acetonitrile in 0.1% aqueous formic acid). The fractions containing the product were combined and lyophilized to give the title compound as a white solid. The yield was 0.032 g.

m/z 674(M+H) ⁺

¹ H NMR (400 MHz, D ₆ -DMSO, 80 ° C): δ 8.15 (s, 2H); 7.88 (s, 1H); 6.87 (d, J = 8.3 Hz, 1H); 6.69 (d, J = 8.3) Hz, 1H); 4.57-4.50 (m, 1H); 3.71-3.58 (m, 8H); 3.36-3.25 (m, 2H); 2.79-2.59 (m, 3H); 2.40-2.31 (m, 2H); 2.32-2.22(m,4H);1.73-1.63(m,2H);1.59-1.48(m,2H);1.44-1.32(m,8H);1.31-1.16(s,10H);0.99-0.93(m , 2H) plus 4 unobserved exchangeables.

Example 281

(R)-4-hydroxy-7-(1-hydroxy-2-(4-(3-(4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diaza) Heterospiro[5.5]undecane-9-yl)propylthio)butylamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

a) 4-(2-(1,3-dioxopenta-2-yl)ethylthio)butan-1-ol

A solution of 4-mercaptobutanol (4.936 g) in dry acetonitrile was cooled to 0 ° C under argon and sodium hydride (60% in mineral oil, 2.044 g). The mixture was stirred at 0&lt;0&gt;C for 1 h then 2-(2-bromoethyl)-1,3-dioxopentane (9.26 g). The reaction was quenched with EtOAc (EtOAc)EtOAcEtOAc The organics were combined and washed with brine (150 mL) dried over sodium sulfate. Purification by chromatography on silica gel eluting with EtOAc (EtOAc) The yield was 7.3 g.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ4.98-4.93 (m, 1H); 4.01-3.93 (m, 2H); 3.89-3.83 (m, 2H); 3.69-3.63 (m, 2H); 2.65- 2.58 (m, 2H); 2.59-2.53 (m, 2H); 1.98-1.91 (m, 2H); 1.73-1.62 (m, 4H) plus an unobservable exchange.

b) (9-(3-(4-Hydroxybutylthio)propyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-iso Propylthiazole-4-yl)methanone

4-(2-(1,3-dioxopenta-2-yl)ethylthio)butan-1-ol (Example 281, step a) (500 mg) was dissolved in 80% of aqueous formic acid Solution (6 mL) and the reaction mixture was stirred at room temperature over the weekend. The mixture was diluted with water (10 mL) and EtOAc (EtOAc (EtOAc)EtOAc. Aldehyde oil. This material was dissolved in anhydrous methanol and (2-isopropyl-thiazol-4-yl)-(1-oxa-4,9-diaza-spiro[5.5]undec-4-yl)-methanone was added. Trifluoroacetate (Example 22, step b) (606 mg) and 3 Molecular sieves. Acetic acid (0.2 mL) was added, and the mixture was stirred under argon for 5 min then cooled to &lt;RTI ID=0.0&gt;&gt; The cooling bath was removed and the reaction mixture was stirred at rt overnight. The mixture was filtered and then applied directly to the SCX-2 column and washed with methanol then the product was dissolved in 2N ammonia methanol. It was further purified by silica gel chromatography eluting with 10% methanol in DCM to give the sub-title compound as light green oil. The yield was 0.37 g.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ7.85 (s, 1H); 4.02-3.87 (m, 2H); 3.80-3.73 (m, 3H); 3.69-3.63 (m, 3H); 3.39-3.23 ( m,5H); 2.86-2.74 (m, 2H); 2.67-2.59 (m, 2H); 2.58-2.50 (m, 5H); 1.96-1.81 (m, 4H); 1.74-1.63 (m, 4H); 1.41 (d, J = 6.9 Hz, 6H).

c) (R)-4-hydroxy-7-(1-hydroxy-2-(4-(3-(4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9- Diazaspiro[5.5]undecane-9-yl)propylthio)butylamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

It was dissolved in anhydrous DCM (2 mL) with chloroform (0.068 mL) and the solution was cooled to -78. DMSO (0.12 mL) in DCM (0.03 mL) was added dropwise and mixture was stirred. After 15 minutes, (9-(3-(4-hydroxybutylthio)propyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl was added dropwise. a solution of (2-isopropylthiazol-4-yl)methanone (Example 281, step b) (290 mg) in DCM (1 mL) and stirring for 15 min then triethylamine (0.44) mL). The cooling bath was removed and the reaction allowed to warm to room temperature. After 1 h, the reaction mixture was crystalljjjjjjjjjjjjjj The layers were separated and the aqueous extracted with DCM (2×5 mL). The organics were combined, washed with brine (5 mL) dried over sodium sulfate and evaporated

The aldehyde (195 mg) and ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, examples 1, step d) (170 mg) was stirred and added in anhydrous methanol (8 mL) Molecular sieves and acetic acid (0.07 mL). After stirring for 5 minutes, the reaction mixture was cooled to 0 ° C and sodium triacetoxyborohydride (200 mg) was added. The cooling bath was removed and the reaction mixture was stirred at room temperature over the weekend. The mixture was filtered and evaporated. Purified by preparative HPLC (Phenomenex Gemini) , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The product-containing fractions were combined and lyophilized to give the title compound as a white solid. The yield was 0.006 g.

m/z 664(M+H) ⁺

¹ H NMR (400 MHz, D ₄ -MeOH): δ 8.48 (s, 1.8H); 7.78 (s, 1H); 6.95 (d, J = 8.3 Hz, 1H); 6.73 (d, J = 8.3 Hz, 1H); 4.97-4.90 (m, 1H); 3.89-3.56 (m, 6H); 3.38-3.24 (m, 1H); 3.13-3.04 (m, 2H); 3.04-2.96 (m, 2H); 2.97- 2.61(m,6H); 2.60-2.49(m,4H); 2.04-1.91(m,2H);1.91-1.70(m,5H);1.70-1.57(m,3H);1.38(d,J=6.9 Hz, 6H) plus 4 unobserved exchangeables.

Example 282

4-hydroxy-7-((1R)-1-hydroxy-2-(4-(3-(4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diaza) Heterospiro[5.5]undecane-9-yl)propylsulfinyl)butylamino)ethyl)benzo[d]thiazole-2(3H)-ketocarboxylate

Trifluoroacetic acid (0.06 mL) was added to (9-(3-(4-hydroxybutylthio)propyl)-1-oxa-4,9-diazaspiro[5.5]undecane- 4-(4-Iso)(2-isopropylthiazol-4-yl)methanone (Example 281, step b) (360 mg) in EtOAc. The solution was cooled to 0 ° C and allowed to stir for 5 min before adding Dess-Martin periodinane (502 mg). The reaction mixture was stirred at room temperature for 1 h then quenched with saturated sodium sulphate (14 mL) and saturated aqueous sodium hydrogen carbonate (14 mL). Ethyl acetate (30 mL) was added, and the mixture was stirred for 5 min. The aqueous layer was extracted with EtOAc (2×30 mL). Acetic acid (0.053 mL) was added to the combined organics which was then dried over sodium sulfate, filtered and evaporated to give the oil of the aldehyde. This material was dissolved in anhydrous methanol (15 mL) along with ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (311 mg) and add 3 Molecular sieves and acetic acid (0.07 mL). After stirring for 5 minutes, the reaction mixture was cooled to 0.degree. C. and sodium triacetoxyborohydride (200 mg) was added and stirred at room temperature overnight. The mixture was filtered and evaporated. Purified by preparative HPLC (Phenomenex , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The fractions containing the product were combined and lyophilized to give the title compound as a white solid. The yield was 0.018 g.

m/z 680(M+H) ⁺

¹ H NMR (400 MHz, D ₄ -MeOH): δ 8.40 (s, 2H); 7.78 (s, 1H); 6.96 (d, J = 8.3 Hz, 1H); 6.73 (d, J = 8.3 Hz, 1H) ); 4.98-4.92 (m, 1H); 3.90-3.56 (m, 6H); 3.36-3.24 (m, 1H); 3.19-3.03 (m, 4H); 3.04-2.67 (m, 10H); 2.15-1.95 (m, 4H); 1.93-1.56 (m, 6H); 1.38 (d, J = 6.9 Hz, 6H) plus 4 unobservable exchangeables.

Example 283

4-hydroxy-7-((1R)-1-indolyl-2-(9-(4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro) [5.5] undecane-9-yl)-2-methylindolyl)ethyl)benzo)[d]thiazole-2(3H)-onecarboxylate

a) 9-bromo-2-methylnonan-1-ol

A solution of diisopropylamine (5.42 mL) in dry THF (30 mL) EtOAc (EtOAc) The solution was stirred at 0 ° C for 30 minutes and then cooled to -78 °C. Ethyl propionate (4.01 mL) was added portionwise, and the solution was stirred at -78 ° C for one hour. 1,7-Dibromoheptane (6.63 mL) was added and stirring was continued at -78 °C for 1 hour, then the cooling bath was removed and the mixture was stirred at room temperature overnight. Saturated aqueous ammonium chloride (30 mL) was added to the reaction then ethyl acetate (30 mL). The layers were separated and the aqueous extracted with ethyl acetate (2×30 mL). The combined organics were washed with EtOAc EtOAc. Purification by gel chromatography eluting with 1 to 5% ethyl acetate in petroleum ether (40-60 ° C

This material (0.84 g) was dissolved in dry diethyl ether (27 mL) and cooled to EtOAc. Diisobutylaluminum hydride (1 M in toluene, 6.7 mL) was added dropwise, and the reaction was stirred at 0 ° C for 1 h 15 min. The reaction was stopped by the addition of a saturated aqueous solution of sodium potassium tartrate, and the resulting mixture was stirred for 15 minutes. Ethyl acetate (60 mL) and water (20 mL) were added and the phases were separated. The aqueous layer was extracted with EtOAc (3×30 mL)EtOAc. Purification by chromatography on silica gel eluting with 0 to 10% ethyl acetate in hexanes to give the subtitle compound as a colourless oil. The yield was 0.23 g.

¹ H NMR (400 MHz, CDCl ₃ ): δ 3.51 (dd, J = 10.5, 5.8 Hz, 1H); 3.46-3.38 (m, 3H); 1.90-1.80 (m, 2H); 1.66-1.55 (m, 2H); 1.47-1.22 (m, 9H); 0.92 (d, J = 6.7 Hz, 3H) plus an unobservable exchangeable substance.

b) (9-(9-Hydroxy-8-methylindolyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-isopropylthiazole -4-yl)methanone

(2-Isopropyl-thiazol-4-yl)-(1-oxa-4,9-diaza-spiro[5.5]undec-4-yl)-methanone di-trifluoroacetate ( Example 22, step b) (500 mg) was applied to a methanol-wet SCX-2 column, washed with methanol, then eluted with 2N ammonia in methanol and evaporated to give a free base (279 mg). This material was dissolved in acetonitrile (10 mL) along with 9-bromo-2-methylnonan-1-ol (Example 283, step

In a) (220 mg), triethylamine (0.223 mL) was added. The reaction mixture was heated to 60 ° C overnight and then concentrated. Purification by silica gel chromatography eluting with 0 to 10% MeOH in DCM The yield was 0.38 g.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ7.87 (s, 1H); 4.06-3.90 (m, 2H); 3.83-3.71 (s, 4H); 3.52-3.37 (m, 4H); 3.05-2.85 ( m, 4H); 2.48-2.32 (m, 2H); 2.15-2.05 (m, 2H); 1.97-1.84 (m, 2H); 1.66-1.50 (m, 4H); 1.48-1.39 (m, 6H); 1.38-1.21 (m, 8H); 0.91 (d, J = 6.7 Hz, 3H) plus an unobservable exchangeable.

c) (9-(9-Bromo-8-methylindolyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-isopropylthiazole -4-yl)methanone

(9-(9-Hydroxy-8-methylindolyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-isopropylthiazole-4 Methyl ketone (Example 283, step b) (100 mg), carbon tetrabromide (143 mg) and imidazole (30 mg) were dissolved in DCM (5 mL). The solution was cooled to 0.degree. C. and a solution of triphenylphosphane (85 mg) in DCM (1 mL). The reaction mixture was stirred at this temperature for 1 hour and then at room temperature for 2 hours. An additional amount of carbon tetrabromide (150 mg) was added and the mixture was cooled to 0.degree. C. and triphenylphosphine (114 mg) was added dropwise in DCM (1 mL). Stir at room temperature for 1 hour, then add water and separate the phases. The organic phase was evaporated and the product was purified eluting eluting elut elut The yield was 0.088 g.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ7.87 (s, 1H); 4.03-3.91 (m, 2H); 3.81-3.70 (m, 4H); 3.43-3.313 (m, 4H); 3.06-2.86 ( m, 4H); 2.49-2.34 (m, 2H); 2.15-2.05 (m, 2H); 1.98-1.86 (m, 2H); 1.57-1.50 (m, 2H); 1.47-1.39 (m, 6H); 1.37-1.19 (m, 10H); 1.00 (d, J = 6.6 Hz, 3H).

d) 4-hydroxy-7-((1R)-1-hydroxy-2-(9-(4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)-2-methyldecylamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

( R )-7-(2-Amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) 263 mg) was dissolved in methanol (10 mL) and sodium methoxide (54 mg) was added and the mixture was stirred for 25 min. The solid was collected by filtration and washed with methanol and dried then evaporated A portion of this material (61 mg) was dissolved in DMF (3.5 mL) and (9-(9-bromo-8-methylindolyl)-1-oxa-4,9-diazaspiro[5.5] Carboxy-4-yl)(2-isopropylthiazol-4-yl)methanone (Example 283, step c) (71 mg) and diisopropylethylamine (0.023 mL) were combined. The reaction mixture was heated to 50 &lt;0&gt;C overnight, then cooled and partitioned between ethyl acetate (10 mL) and water (10 mL). The organics were dried over sodium sulfate, filtered and concentrated. Purified by preparative HPLC (Phenomenex , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The fractions containing the product were combined and lyophilized to give the title compound as a white solid. The yield was 0.002 g.

m/z 674(M+H) ⁺

¹ H NMR (400 MHz, D ₄ -MeOH): δ 8.51 (s, 1H); 7.78 (s, 1H); 6.92 (d, J = 8.3 Hz, 1H); 6.70 (d, J = 8.3 Hz, 1H) ); 3.85-3.55 (s, 6H); 2.92-2.80 (m, 2H); 2.76-2.34 (m, 8H); 1.97-1.82 (m, 2H); 1.77-1.44 (m, 5H); 1.37 (d) , J=6.9 Hz, 6H); 1.36-1.17 (m, 9H); 1.17-1.04 (m, 1H); 0.92 (dd, J=6.6, 3.2 Hz, 3H) plus two signals obscured by solvent and Four unobserved exchangeables.

Example 284 (R)-4-hydroxy-7-(1-hydroxy-2-(7-(4-(5-isopropylthiophen-3-carbonyl)-1-oxa-4,9-diazaspiro[ 5.5] undecane-9-yl)heptylamino)ethyl)benzo[d]thiazole-2(3H)-one formate

a) (9-(7-Hydroxyheptyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(5-isopropylthiophen-3-yl) Ketone

A solution of 7-bromo-1-heptanol (380 mg) in acetonitrile (3.9 mL) was added to (5-isopropylthiophen-3-yl)(1-oxa-4,9-diazaspiro[5.5 a solution of the undecane-4-yl)methanone (Example 275, step i) (400 mg) and triethylamine (0.361 mL) in EtOAc (EtOAc) The resulting mixture was stirred at 60 ° C for 16 hours. The solvent was removed in vacuo and EtOAc EtOAc m. Purification by silica gel chromatography eluting with 0 to 10% MeOH in DCM The yield was 0.305 g.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ7.26 (s, 1H); 6.84 (s, 1H); 3.74-3.67 (m, 4H); 3.63 (t, J = 6.4Hz, 2H); 3.57-3.48 (m, 2H); 3.35-3.10 (m, 2H); 2.99-2.70 (m, 3H); 2.28-1.98 (m, 4H); 1.92-1.74 (m, 2H); 1.67-1.47 (m, 4H) ; 1.41-1.34 (m, 5H); 1.34 (d, J = 6.9 Hz, 6H); 1.28-1.23 (m, 1H) plus an unobservable exchangeable.

b) (R)-4-hydroxy-7-(1-hydroxy-2-(7-(-4-(5-isopropylthiophene-3-carbonyl)-1-oxa-4,9-diaza Hetero[5.5]undecane-9-yl)heptylamino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

Adding trifluoroacetic acid (0.054 mL) to (9-(7-hydroxyheptyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl) (5-iso Propylthiophen-3-yl)methanone (Example 284, step a) (295 mg) in EtOAc (EtOAc) The mixture was stirred for 5 minutes, then Dess-Martin periodinane (503 mg) was added and stirred for 1 hour and 50 minutes. The reaction mixture was quenched by the addition of saturated aqueous sodium thiosulfate (4 mL) and saturated aqueous sodium hydrogen carbonate (4 mL). Ethyl acetate (25 mL) was added and the mixture was stirred vigorously for 5 min then the phases were separated. The aqueous layer was extracted with EtOAc (5 mL). The organics were combined, washed with brine (5 mL) EtOAc (EtOAc)EtOAc. This material was dissolved in anhydrous methanol (5 mL), followed by the addition of ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride (WO 2007027134, Example 1, step d) (202 mg), and the mixture was stirred for 5 minutes. The reaction mixture was cooled to 0 ° C, sodium cyanoborohydride (66 mg) was added and stirred at 0&lt;0&gt;C for 2 s. The mixture was concentrated in vacuo and partitioned between EtOAc (EtOAc) (EtOAc) The layers were separated and the organic phase was dried with sodium sulfate, filtered and evaporated. Purified by preparative HPLC (Phenomenex , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The fractions containing the product were combined and lyophilized to give the title compound as a white solid. The yield was 0.037 g.

m/z 631(M+H) ⁺

¹ H NMR (400 MHz, D _{4 -} MeOH): δ 8.49 (s, 2H); 7.45 (s, 1H); 6.95 (d, J = 8.3 Hz, 1H); 6.91 (s, 1H); , J = 8.3 Hz, 1H); 4.93 (dd, J = 9.1, 4.5 Hz, 1H); 3.79-3.47 (m, 6H); 3.20-3.04 (m, 3H); 3.01-2.94 (m, 3H); 2.92-2.70(m,4H);2.08-1.97(m,2H);1.75-1.56(m,6H);1.43-1.33(m,6H);1.30(d,J=6.9Hz,6H) plus one factor A solvent-blurred signal and four unobserved exchangeables.

Example 285 (R)-4-hydroxy-7-(1-hydroxy-2-(9-(4-(2-isopropylthiazol-4-carbonyl)-1-oxa-4,9-diazaspiro 15.51 Undecane-9-yl)-4,4-dimethylmercapto)ethyl)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

a) 2-(5-Bromopentyloxy)tetrahydro-2H-pyran

To a solution of 5-bromopentanol (10 g) in DCM (150 mL) was added &lt;RTI ID=0.0&gt;&gt; The temperature was stirred under nitrogen for 21 hours. The solution was diluted with diethyl ether then washed with dilute brine (1:1 brine: water: EtOAc) Purification by silica gel chromatography, eluting with 0 to 50% ethyl acetate in hexanes to give the subtitle compound as a colourless liquid. The yield was 14.14 g.

¹ H NMR (400 MHz, CDCl ₃ ): δ 4.59 - 4.56 (m, 1H); 3.91-3.82 (m, 1H); 3.79-3.71 (m, 1H); 3.57-3.47 (m, 1H); 3.36 (m, 3H); 1.94-1.76 (m, 3H); 1.76-1.66 (m, 1H); 1.65-1.46 (m, 8H).

b) 2,2-Dimethyl-7-(tetrahydro-2H-piperidin-2-yloxy)heptanoic acid ethyl ester

A solution of diisopropylamine (1.78 mL) in dry EtOAc (EtOAc) (EtOAc) Stir at 0 ° C for 30 minutes, then cool to -78 ° C and treat with ethyl isobutylate (1.55 mL). The solution was stirred at -78 °C for 45 min then added 2-(5-bromopentyloxy)tetrahydro-2H-pyran (Example 285, step a) (3.20 g) in dry THF (1 mL) Solution. The reaction mixture was stirred at -78 °C for 10 minutes and then allowed to warm to room temperature overnight. The reaction mixture was cooled to EtOAc (3 mL)EtOAc The aqueous phase was extracted with EtOAc (EtOAc) (EtOAc)EtOAc. Purification by chromatography on silica gel eluting with EtOAc EtOAc (EtOAc) The yield was 2.22 g.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ4.59-4.55 (m, 1H); 4.11 (q, J = 7.1Hz, 2H); 3.90-3.82 (m, 1H); 3.72 (dt, J = 9.6, 6.8 Hz, 1H); 3.53-3.45 (m, 1H); 3.37 (dt, J = 9.6, 6.6 Hz, 1H); 1.88-1.77 (m, 1H); 1.75-1.67 (m, 1H); 1.64-1.48 (m, 8H); 1.40-1.30 (m, 2H); 1.29-1.20 (m, 5H); 1.15 (s, 6H).

c) 2,2-dimethyl-7-(tetrahydro-2H-piperidin-2-yloxy)heptan-1-ol

A solution of ethyl 2,2-dimethyl-7-(tetrahydro-2H-piperidin-2-yloxy)heptanoate (Example 285, step b) (2.22 g) Treated with diisobutylaluminum hydride (1 M in toluene, 17.1 mL) at 0 ° C under nitrogen for 5 min. After 1 hour at 0 ° C, the reaction mixture was carefully treated with aqueous sodium potassium tartite, diluted with diethyl ether, and the mixture was stirred vigorously for 1.5 hours. The phases were separated and the aqueous phase was extracted with diethyl ether (x 2). The combined organic phases were washed with water and brine, dried over magnesium sulfate, filtered and evaporated. Purification by chromatography on silica gel eluting with EtOAc (EtOAc:EtOAc) The yield was 1.83 g.

¹ H NMR (400 MHz, CDCl ₃ ): δ 4.58-4.55 (m, 1H); 3.91-3.83 (m, 1H); 3.77-3.69 (m, 1H); 3.53-3.46 (m, 1H); Dt, J = 9.6, 6.6 Hz, 1H); 3.31 (d, J = 4.1 Hz, 2H); 1.90-1.78 (m, 1H); 1.76-1.64 (m, 1H); 1.64-1.48 (m, 4H) ; 1.40-1.20 (m, 8H); 0.86 (s, 6H) plus an unobserved exchangeable substance.

d) (E)-4,4-Dimethyl-9-(tetrahydro-2H-piperidin-2-yloxy)indole-2-enoate

A solution of chloroform (0.78 mL) in EtOAc (EtOAc) (EtOAc) After 15 minutes, 2,2-dimethyl-7-(tetrahydro-2H-piperidin-2-yloxy)heptan-1-ol was added dropwise (Example 285, step c) (1.73 g) The solution in DCM (10 mL) was stirred for 15 min then triethylamine (4.93 mL). The cooling bath was removed and the reaction allowed to warm to room temperature. After 1 hour, the reaction mixture was treated with saturated aqueous ammonium chloride and layered. The aqueous phase was extracted with DCM (x2) and combined organics washed with brine, dried over magnesium sulfate, filtered and evaporated This material was taken up in DCM, washed with water, dried over magnesium sulfate, filtered and evaporated to give the aldehyde as a colorless oil.

A solution of triethylphosphonium sulphate (1.62 mL) in acetonitrile (8 mL) was treated with lithium chloride (0.43 g) and then cooled to 0 ° C, dropwise in the form of a solution in acetonitrile (2.5 mL) Triethylamine (1.23 mL) was added. The reaction mixture was stirred at 0 ° C for 15 min then a solution of &lt The mixture was allowed to warm to room temperature and stirred for 3 hours, then treated with saturated aqueous ammonium chloride, diethyl ether and water. The layers were separated and the aqueous extracted with EtOAc (EtOAc)EtOAc. Purification by chromatography on silica gel eluting with EtOAc (EtOAc:EtOAc) The yield was 0.89 g.

¹ H NMR (400 MHz, CDCl ₃ ): δ 6.91 (d, J = 15.9 Hz, 1H); 5.71 (dd, J = 15.9, 1.9 Hz, 1H); 4.59 - 4.55 (m, 1H); 4.19 (q) , J=7.1 Hz, 2H); 3.90-3.82 (m, 1H); 3.76-3.68 (m, 1H); 3.53-3.46 (m, 1H); 3.40-3.33 (m, 1H); 1.89-1.77 (m) , 1H); 1.76-1.66 (m, 1H); 1.64-1.48 (m, 6H); 1.39-1.19 (m, 9H); 1.05-1.03 (m, 6H).

e) 4,4-Dimethyl-9-(tetrahydro-2H-piperidin-2-yloxy)decanoic acid ethyl ester

(E)-4,4-Dimethyl-9-(tetrahydro-2H-piperidin-2-yloxy)indol-2-enoate (Example 285, step d) (0.89 g) A solution of IMS (industrial grade in methanol with ethanol) (10 mL) was treated with palladium on carbon (spice tip) under nitrogen, and then the flask was purged with hydrogen (x3) and then stirred overnight under hydrogen atmosphere. An additional portion of the palladium on carbon was added and the hydrogenation was restarted for an additional 24 hours. The catalyst was filtered off through celite and the filtrate was evaporated to give a sub-title compound as a colorless oil. The yield was 0.86 g.

¹ H NMR (400 MHz, CDCl ₃ ): δ 4.59 - 4.56 (m, 1H); 4.12 (q, J = 7.1 Hz, 2H); 3.90-3.83 (m, 1H); 3.75-3.68 (m, 1H) ;3.53-3.46(m,1H);3.41-3.34(m,1H); 2.26-2.19(m,2H);1.88-1.79(m,2H);1.76-1.66(m,2H);1.66-1.43( m, 6H); 1.40-1.14 (m, 9H); 0.84 (s, 6H).

f) ethyl 9-hydroxy-4,4-dimethyl decanoate

Addition of 4,4-dimethyl-9-(tetrahydro-2H-piperidin-2-yloxy)decanoate (Example 285, step e) (0.86 g) in methanol (10 mL) p-Toluenesulfonic acid monohydrate (52 mg), and the reaction mixture was stirred at room temperature for 2 hr. The mixture was diluted with ethyl acetate and water and the phases were separated. The aqueous layer was extracted with EtOAc (EtOAc) (EtOAc)EtOAc. Purification by chromatography on silica gel eluting with EtOAc (EtOAc:EtOAc) The yield was 0.44 g.

_{^{1 H NMR (400MHz, CDCl 3}} ): δ4.12 (q, J = 7.1Hz, 2H); 3.67-3.61 (t, J = 6.4Hz, 2H); 2.28-2.20 (m, 2H); 1.61-1.50 (m, 4H); 1.49-1.37 (m, 1H); 1.38-1.22 (m, 7H); 1.21-1.12 (m, 2H); 0.85 (s, 6H).

g) ethyl 9-bromo-4,4-dimethyl decanoate

A solution of 9-hydroxy-4,4-dimethyl decanoate (Example 285, step f) (0.44 g) in DCM (19 mL) Treatment with phenylphosphine (0.60 g) and the reaction mixture was stirred at room temperature overnight. Additional amounts of carbon tetrabromide (0.76 g) and triphenylphosphine (0.60 g) were added and stirred for 3 hours. The solvent was evaporated, then the mixture was triturated with cyclohexane and filtered to afford a solid. The solid was washed with cyclohexane and the filtrate was concentrated to ca. 15 mL and allowed to stand overnight. The solid which precipitated from the solution was filtered off, washed with hexanes and evaporated to give a pale brown oil. The yield was 0.56 g.

_{^{1 H NMR (300MHz, CDCl 3}} ): δ4.12 (q, J = 7.1Hz, 2H); 3.41 (t, J = 6.8Hz, 2H); 2.28-2.20 (m, 2H); 1.93-1.80 (m , 2H); 1.59-1.49 (m, 2H); 1.46-1.34 (m, 2H); 1.32-1.21 (m, 5H); 1.22-1.13 (m, 2H); 0.85 (s, 6H).

h) 9-bromo-4,4-dimethylnonan-1-ol

A solution of ethyl 9-bromo-4,4-dimethyl decanoate (Example 285, step g) (0.56 g) in dry diethyl ether (19 mL). (1 M in toluene, 4.20 mL), and the mixture was stirred at 0 ° C for 1 hour and 15 min. The reaction mixture was carefully treated with aqueous sodium potassium tartrate solution and stirred for 1 hour. The phases were separated and the aqueous layer was evaporated (jjjjjjjli The yield was 0.42 g.

¹ H NMR (400 MHz, CDCl ₃ ): δ 3.62 (t, J = 6.7 Hz, 2H); 3.41 (t, J = 6.9 Hz, 2H); 1.91-1.82 (m, 2H); 1.55-1.45 (m) , 2H); 1.46-1.35 (m, 2H); 1.30-1.15 (m, 6H); 0.85 (s, 6H) plus an unobservable exchangeable substance.

i) (9-(9-Hydroxy-6,6-dimethylindolyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-iso Propylthiazole-4-yl)methanone

(2-isopropyl-thiazol-4-yl)-(1-oxa-4,9-diaza-spiro[5.5]undec-4-yl)-methanone di-trifluoroacetate (implementation Example 22, step b) (0.40 g) was applied to a methanol-wet SCX-2 cartridge eluted with methanol then eluted with 2N ammonia in methanol and evaporated to yield the free base (227 mg). This was treated with 9-bromo-4,4-dimethylnonan-1-ol (Example 285, step h) (0.11 g) in acetonitrile ( 8.8 mL) followed by triethylamine (0.20 mL) . The mixture was heated at 60 ° C for 21 h before the solvent was evaporated. Purification by chromatography on silica gel eluting with EtOAc EtOAc EtOAc EtOAc The yield was 0.295 g.

¹ H NMR (400 MHz, D ₆ -DMSO): δ 8.00 (s, 1H); 4.35 - 4.30 (m, 1H); 3.76 - 3.45 (m, 6H); 3.37 - 3.28 (m, 1H); 2.03 (m, 6H); 1.72-1.58 (m, 2H); 1.57-1.44 (m, 2H); 1.43-1.28 (m, 10H); 1.27-1.05 (m, 9H); 0.81 (s, 6H) plus An unobserved exchangeable substance.

j) (R)-4-hydroxy-7-(1-hydroxy-2-(9-(4-(2-isopropylthiazole-4-carbonyl)-1-oxa-4,9-diaza Spiro[5.5]undecane-9-yl)-4,4-dimethylhydrazino)ethyl)benzo[d]thiazole-2(3H)-onecarboxylate

(9-(9-Hydroxy-6,6-dimethylindolyl)-1-oxa-4,9-diazaspiro[5.5]undecane-4-yl)(2-isopropyl A solution of thiazol-4-yl)methanone (Example 285, step i) (0.255 g) in DCM (5.3 mL) Add Dess-Martin periodinane (0.339 g). The solution was stirred at room temperature for 1 hour, after which an additional amount of Dess-Martin periodinane (0.113 g) was added and stirring was continued for 30 minutes. The reaction mixture was treated with saturated aqueous sodium thiosulfate and saturated aqueous sodium hydrogen carbonate (1:1), diluted with DCM and stirred for 10 min. Ethyl acetate was added and the phases were separated. The aqueous layer was extracted with ethyl acetate (×2). The combined organics were washed with a saturated aqueous solution of sodium bicarbonate and then dried over magnesium sulfate, and then acetic acid (0.046 <RTIgt; This was suspended in anhydrous methanol (9 mL) with ( R )-7-(2-amino-1-hydroxyethyl)-4-hydroxybenzo[d]thiazole-2(3H)-one hydrochloride ( WO 2007027134, Example 1, step d) (0.210 g), 3 The molecular sieves were treated with acetic acid (0.046 mL) and the mixture was stirred for 5 min. The reaction mixture was cooled to 0.degree. C. and sodium triacetoxyborohydride (113 mg) was added and stirred at room temperature for 24 hours. The mixture was filtered and concentrated under vacuum. Purified by gel chromatography to dissolve in 5, 6, 7, 8, 9, 10 and 12% (10% aqueous ammonia solution in methanol) in DCM, followed by preparative HPLC (Phenomenex) , Gradient: 5 to 40% acetonitrile in 0.1% aqueous formic acid). The fractions containing the product were combined and lyophilized to give the title compound as a white solid. The yield was 0.024 g.

m/z 688(M+H) ⁺

¹ H NMR (400 MHz, D ₆ -DMSO, 80 ° C): δ 8.26 - 8.13 (m, 1H); 7.89 (s, 1H); 6.87 (d, J = 8.3 Hz, 1H); 6.69 (d, J = 8.3 Hz, 1H); 4.61-4.53 (m, 1H); 3.69-3.63 (m, 6H); 3.37-3.24 (m, 1H); 2.76-2.66 (m, 2H); 2.59-2.50 (m, 2H) ); 2.40-2.31 (m, 2H); 2.31-2.20 (m, 4H); 1.72-1.63 (m, 2H); 1.58-1.47 (m, 2H); i.44-1.29 (m, 10H); -1.09 (m, 8H); 0.80 (s, 6H) plus four unobservable exchangeables.

The compounds of the invention may be tested for pharmaceutical activity using techniques known in the art, such as, for example:

Adrenergic β2-mediated detection of cAMP production Cell preparation

H292 cells were cultured in RPMI medium containing 10% (v/v) FBS (fetal calf serum) and 2 mM L-glutamine in a 225 cm ₂ flask incubator at 37 ° C under 5% CO ₂ .

experimental method

Whereby Accutase ^TM cell detachment solution for 15 minutes to remove the adhesion H292 cells obtained from tissue culture flasks. The flask was incubated for 15 minutes at 37 ° C, 5% CO ₂ in a humidified incubator. The detached cells were resuspended in RPMI medium (containing 10% (v/v) FBS and 2 mM L-glutamine) at 0.1 × 10 ⁶ cells/mL. 100 μL of 10000 cells were added to each well of a tissue culture-treated 96-well plate, and the cells were incubated overnight at 37 ° C, 5% CO ₂ in a humidified incubator. The medium was removed, the cells were washed twice with 100 μL of assay buffer, and replaced with 50 μL of assay buffer (HBSS solution containing 10 mm HEPES pH 7.4 and 5 mM glucose). The cells were allowed to rest at room temperature for 20 minutes, after which 25 μL of rolipram (1.2 mM in a detection buffer containing 2.4% (v/v) dimethyl hydrazine) was added. The cells were incubated with rolipram for 10 minutes, after which test compounds were added and the cells were incubated for 60 minutes at room temperature. The final concentration of rolipram in the assay was 300 μM and the final adjuvant concentration was 1% (v/v) dimethyl sulfoxide. The reaction was stopped by removing the supernatant, washed once with 100 μL of detection buffer and replaced with 50 μL of lysis buffer. The cell monolayer was frozen at -80 °C for 30 minutes (or overnight).

A1phaScreen ^TM cAMP detection

Determination of cAMP (cyclic adenosine monophosphate) concentration of cell-based methods using AlphaScreen ^TM lysate. The frozen cell plates were thawed on a plate shaker for 20 minutes, after which 10 μL of cell lysate was transferred to a 96-well white plate. The biotinylated cAMP 40μL pre-incubated beads of mixed AlphaScreen ^TM Detection was added to each well, the plates incubated at room temperature in the dark for 3 hours. AlphaScreen ^TM-based signal having the spectrum splitter EnVision (Perkin-Elmer Inc.) recommended by the manufacturer of the set measurement. The cAMP concentration was determined with reference to a calibration curve measured using the standard cAMP concentration in the same experiment. The concentration response curve was constructed for the agonist, and the data was fitted to a four-parameter logistic equation to determine the pEC ₅₀ and intrinsic activity. The intrinsic activity is expressed as the ratio of the maximum activity of formoterol measured in each experiment.

Combination of muscarinic receptors

The affinity of the compound for binding to the M ₃ receptor (pIC ₅₀ ) is based on the proximity scintillation detection (SPA) mode, and the expression of human muscarinic acetylcholine M by [ ³ H]N-methyl decylamine (NMS) _{3 The} receptor (M ₃ -ACh) CHO-Kl (Chinese big hamster ovary) cell membrane competitive binding determination.

The SPA beads were pre-coated with a film, and then the compound of the invention, [ ³ H]NMS 0.1 nM, quarter Kd (experimental dissociation constant) and detection buffer (containing 5 mM) were serially diluted at 2 Mg beads/well. Incubation of MgCl ₂ with 20 mM HEPES pH 7.4). The assay was performed in the presence of 1% (v/v) dimethyl hydrazine (DMSO) at a final volume of 200 μL. Total binding ^[3 H] NMS of competing compound based on a decision absent, and ^[3 H] NMS based non-specific binding in the 1μM Atropine (atropine) the determined presence. The plate was incubated at room temperature for 16 hours and read using a normalized ³ H methods follow on Wallac Microβ ^TM. It was determined that pIC ₅₀ - is defined as a negative logarithm of the molar concentration of the desired compound minus 50% specificity [ ³ H]-NMS binding.

The compounds of the invention were tested according to the aforementioned tests and the following results were obtained:

The β-2 effect % of the compounds of Examples 116 to 263 at 1 μM was measured using the following experimental method.

Adrenaline β2-mediated cAMP production Cell preparation

H292 cells were cultured in RPMI medium containing 10% (v/v) FBS (fetal calf serum) and 2 mM L-glutamine in a 225 cm ₂ flask incubator at 37 ° C under 5% CO ₂ .

experimental method

Compounds were prepared by first dilution in DMSO to produce a 1 mM stock concentration. Thereafter, a 1:10 dilution of the stock concentration (10 μl of the starting compound + 90 μl of DMSO) was followed to yield a 0.1 mM compound.

Compound addition plates were prepared by further 1:25 dilution in a detection buffer containing 4% dimethyl hydrazine (HBSS solution containing 10 mM HEPES pH 7.4 and 5 mM glucose). The final compound concentration in the assay was 1 μM.

Whereby Accutase ^TM cell detachment solution for 15 minutes to remove the adhesion H292 cells obtained from tissue culture flasks. The flask was incubated for 15 minutes at 37 ° C, 5% CO ₂ in a humidified incubator. The detached cells were resuspended in RPMI medium (containing 10% (v/v) FBS and 2 mM L-glutamine) at 0.1 × 10 ⁶ cells/mL. 100 μL of 10000 cells were added to each well of a tissue culture-treated 96-well plate, and the cells were incubated overnight at 37 ° C, 5% CO ₂ in a humidified incubator. The medium was removed, the cells were washed twice with 100 μL of assay buffer, and replaced with 50 μL of assay buffer (HBSS solution containing 10 mm HEPES pH 7.4 and 5 mM glucose). The cells were allowed to rest at room temperature for 20 minutes, after which 25 μL of rolipram (1.2 mM, composed of assay buffer) was added. The cells were incubated with rolipram for 10 minutes, after which 25 μL of the test compound was added and the cells were incubated at room temperature for 60 minutes. The final concentration of rolipram in the assay was 300 μM and the final adjuvant concentration was 1% (v/v) dimethyl sulfoxide. The reaction was stopped by removing the supernatant, washed once with 100 μL of detection buffer and replaced with 50 μL of lysis buffer. The cell monolayer was frozen at -80 °C for 30 minutes (or overnight).

A1phaScreen ^TM cAMP detection

Determination of cAMP (cyclic adenosine monophosphate) concentration of cell-based methods using AlphaScreen ^TM lysate. The frozen cell plates were thawed on a plate shaker for 20 minutes, after which 10 μL of cell lysate was transferred to a 96-well white plate. The mixed AlphaScreen ^TM 40μL detection beads (containing equal volumes of donor beads (biotinylated cAMP pre-incubated in the dark for 30 minutes) and the receptors beads) was added to each well, the plate in the dark at room Incubate for 3 hours. AlphaScreen ^TM-based signal having the spectrum splitter EnVision (Perkin-Elmer Inc.) recommended by the manufacturer of the set measurement. The cAMP concentration produced per well was calculated by reference to the determined cAMP standard curve as a percentage of the maximum response produced by 1E-07M formoterol as determined by the same experiment.

The M3 inhibition % of the compounds of Examples 116 to 263 at 1 μM was measured using the following experimental method.

Combination of muscarinic receptors

The activity of the compound for the M ₃ receptor (% of specific binding inhibition) is in the proximity scintillation detection (SPA) mode, and the human muscarinic choline is represented by [ ³ H]N-methyl decylamine (NMS). The competitive binding of the CHO-Kl (Chinese big hamster ovary) cell membrane of the M ₃ receptor (M ₃ -ACh).

SPA beads were pre-coated with 1 μM of the compound of the invention, [ ³ H]NMS 0.1 nM, quarter Kd (experimental dissociation constant) and detection buffer (containing 5 mM MgCl) at 2 mg beads/well Incubate with ₂ mM HEPES pH 7.4). The assay was performed in the presence of 1% (v/v) dimethyl hydrazine (DMSO) at a final volume of 200 μL. Total binding ^[3 H] NMS of competing compound based on a decision absent, and ^[3 H] NMS based non-specific binding in the 1μM Atropine (atropine) the determined presence. The plate was incubated at room temperature for 16 hours and read using a normalized ³ H methods follow on Wallac Microβ ^TM. The activity of the compound measured at 1 μM was defined as the % inhibition of specific [ ³ H]-NMS binding.

Claims (14)

a compound of formula I Where Ar is selected from any of the following Wherein M ¹ is S, C(O), NH, CH ₂ , CH ₂ CH ₂ , CH=CH, CH ₂ O or OCH ₂ ; M ² is S, C(O), NH, CH ₂ , CH ₂ CH ₂ , CH=CH, CH ₂ O or OCH ₂ ; A ³ may also be CH ₂ OH, NHCHO, NHS(O) ₂ NA ¹⁵ A ¹⁶ or NHSO ₂ A ¹⁷ ; A ¹⁵ or A ^{16 is} independently hydrogen or C _1-6 alkyl; and A ¹⁷ is C _1-6 alkyl; L represents a linker, a straight or branched hydrocarbon group containing up to 10 carbon atoms; wherein up to three carbon atoms in the chain are optionally Substituted by up to four substituents independently selected from the group consisting of: halogen, S(O) _0-2 R ⁵⁶ , NR ⁵⁷ R ⁵⁸ , S(O) ₂ NR ⁵⁹ R ⁶⁰ , C(O)NR ⁶¹ R ⁶² , C(O)OR ⁶³ , NR ⁶⁴ S(O) ₂ R ⁶⁵ , NR ⁶⁶ C(O)R ⁶⁷ , NR ⁶⁸ C(O)OR ⁶⁹ , NR ⁷⁰ C(O)NR ⁷¹ R ⁷² , OR ⁷³ , C _{a 1-6} alkyl group and a C _3-6 cycloalkyl group, wherein the C _1-6 alkyl group and the C _3-6 cycloalkyl group are optionally substituted with up to three substituents independently selected from the group consisting of halogen, hydroxy and C _{1 -6} alkoxy; wherein up to five carbon atoms in the chain may be replaced by groups independently selected from: O, NR ⁴⁵ , S, S(O), S(O) ₂ , C(O)O, OC(O), NR ⁴⁶ C(O), C(O)NR ⁴⁷ , NR ⁴⁸ S(O) ₂ , S(O) ₂ NR ⁴⁹ , NR ⁵⁰ C( O) NR ⁵¹ , NR ⁵² S(O) ₂ NR ⁵³ , OC(O)NR ⁵⁴ , NR ⁵⁵ C(O)O, with the proviso that any of the groups in the chain are separated by at least 2 carbon atoms And up to six carbon atoms in its mid-chain can form part of a monocyclic or bicyclic aliphatic, heteroaliphatic, aromatic or heteroaromatic ring having up to four heteroatoms independently selected from N, O or S The ring contains up to 10 ring atoms, wherein the ring is optionally substituted with up to three substituents independently selected from the group consisting of: halogen, S(O) _0-2 R ⁵⁶ , NR ⁵⁷ R ⁵⁸ , S(O) ₂ NR ⁵⁹ R ⁶⁰ , C(O)NR ⁶¹ R ⁶² , C(O)OR ⁶³ , NR ⁶⁴ S(O) ₂ R ⁶⁵ , NR ⁶⁶ C(O)R ⁶⁷ , NR ⁶⁸ C(O)OR ⁶⁹ , NR ⁷⁰ C(O)NR ⁷¹ R ⁷² , OR ⁷³ , C _1-6 alkyl and C _3-6 cycloalkyl, wherein C _1-6 alkyl and C _3-6 cycloalkyl are optionally up to three independent Substituted with a substituent selected from the group consisting of halogen, hydroxy and C _1-6 alkoxy; the chain may comprise up to three independently selected such rings; wherein R ⁵⁶ , R ⁶⁵ and R ⁶⁹ each independently represent C _{1- alkyl} or C _3-6 cycloalkyl wherein the C _1-6 alkyl and C _3-6 cycloalkyl may optionally be substituted with up to three substituents independently selected from the substituents: , Hydroxy, C _1-6 alkoxy; and ^{R 45, R 46, R 47} , R 48, R 49, R 50, R 51, R 52, R 53, R 54, R 55, R 57, R ⁵⁸ , R ⁵⁹ , R ⁶⁰ , R ⁶¹ , R ⁶² , R ⁶³ , R ⁶⁴ , R ⁶⁶ , R ⁶⁷ , R ⁶⁸ , R ⁷⁰ , R ⁷¹ , R ⁷² and R ⁷³ each independently represent hydrogen, C _1-6 alkane Or a C _3-6 cycloalkyl group, wherein the C _1-6 alkyl group and the C _3-6 cycloalkyl group are optionally substituted with up to three substituents independently selected from the group consisting of halogen, hydroxy, C _1-6 alkane Any one of R ⁵⁷ and R ⁵⁸ , R ⁵⁹ and R ⁶⁰ , R ⁶¹ and R ⁶² , or R ⁷¹ and R ⁷² together with the nitrogen atom linking the two may form a 4 to 8 member aliphatic heterocyclic ring. Wherein the heterocyclic ring may comprise up to three heteroatoms independently selected from N, O and S, wherein the ring may optionally be substituted with up to three substituents independently selected from the group consisting of halogen, hydroxy, _C1-6 alkyl Or a C _3-6 cycloalkyl group, wherein the C _1-6 alkyl group and the C _3-6 cycloalkyl group are optionally substituted with up to three substituents independently selected from the group consisting of halogen, hydroxy and C _1-6 alkoxy And the chain thereof may additionally comprise up to three carbon-carbon double bonds; wherein the chain may additionally comprise up to Carbon - carbon triple bond; L ¹ and L ² independently represent hydrogen, C _1-6 alkyl or C _3-6 cycloalkyl; L ³ and L ⁴ independently represent hydrogen, C _1-6 alkyl or C _{a 3-6} cycloalkyl group, wherein the C _1-6 alkyl group and the C _3-6 cycloalkyl group are optionally substituted with up to three substituents independently selected from the group consisting of halogen, hydroxy and C _1-6 alkoxy; Further, L ¹ and/or L ³ may be bonded to a carbon atom of a hydrocarbyl chain in the linker L to form an aliphatic ring having up to 6 ring atoms, wherein each ring may comprise up to three independently selected from N, O and a hetero atom of S; R ¹ represents a C _1-6 alkyl group, a C _3-6 cycloalkyl group, a phenyl 5 to 6 membered heteroaryl ring or having up to 10 atoms and having up to three independently selected from N, O Or a fused aromatic or fused heteroaryl ring of a hetero atom of S; when R ¹ is a cycloalkyl, benzene or heteroaryl ring, it may optionally be substituted with up to three substituents independently selected from the group consisting of C: (O)OR ¹² , OR ²² , C _1-7 alkyl (which may optionally be substituted by up to six substituents independently selected from halogen), C _3-8 cycloalkyl, benzene ring, 4 to 8 member aliphatic ring and 5-6 heteroaromatic ring, each having up to four independently selected from N, O or S heteroatoms of; when R ¹ is condensed When the aromatic or fused heteroaryl ring, which may optionally be substituted with up to three independently selected from halogen and C _1-6 alkyl The substituents; wherein one of ^a C _1-6 alkyl group or two carbon chains R The atom may be replaced by O, S or N; and when R ¹ is C _1-6 alkyl, it may optionally be up to three independently selected from the group consisting of benzene rings and each having up to four independently selected from N, O or S. Substituents of 4 to 8 membered heteroaliphatic rings and 5 to 6 membered heteroaryl rings of a hetero atom, and each of which is optionally independently selected from the group consisting of OR ²² , C _1-7 alkyl, benzene ring and each Substituent substitution of up to four 4 to 8 membered heteroaliphatic rings and 5 to 6 membered heteroaryl rings independently selected from N, O or S heteroatoms, and when R ¹ is 4 to 8 membered heterolipids When a C ring is substituted with a C _1-6 alkyl group, the ring may be further substituted with a pendant oxy group; or the C _1-6 alkyl chain may be up to 10 atoms and having up to four independently selected from N, O Or a fused aliphatic, fused heteroaliphatic, fused aromatic or fused heteroaryl ring of a hetero atom of S, each ring optionally having up to three independently selected from halogen and C _1-6 alkyl ( It may optionally take up to three substituents independently selected from halogens) And when R ¹ is fused with time of hetero aliphatic ring substituted C _1-6 alkyl, which ring may be further substituted with oxo; R ¹² and R ²² each independently represents hydrogen or C _1-6 alkyl; X represents O; m = 0, 1, 2 or 3; n = 1, 2, 3 or 4; the constraint is that m + n is greater than or equal to 2 W represents CR ²⁷ R ²⁸ CR ²⁹ R ³⁰ ; V is a bond; Z represents a bond, CR ³⁷ R ³⁸ or CR ³⁹ R ⁴⁰ CR ⁴¹ R ⁴² ; Y represents CO, CONR ⁴³ , SO ₂ or SO ₂ NR ⁴⁴ ; R ²⁷ , R ²⁸ , R ²⁹ , R ³⁰ R ³⁷ , R ³⁸ , R ³⁹ , R ⁴⁰ , R ⁴¹ and R ⁴² each independently represent hydrogen, methyl or fluoro; R ⁴³ and R ⁴⁴ each independently represent hydrogen or methyl; or a pharmaceutically acceptable salt thereof. A compound of the formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof, which is 7-(2-(2-fluoro-5-((4-(2-isopropylthiazol-4-carbonyl))- 1-oxa-4,9-diazaspiro[5.5]undecal-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzo[d] Thiazole-2(3H)-one. A compound of the formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof, which is ( R )-7-(2-(2-fluoro-5-((4-(2-isopropyl-2-thiazole-4) -carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzene And [d] thiazole-2(3H)-one di-trifluoroacetate. A compound of the formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof, which is ( R )-7-(2-(2-fluoro-5-((4-(2-isopropyl-2-thiazole-4) -carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzene And [d] thiazole-2(3H)-ketobis(1S)-(+)-10-camphorsulfonate. A compound of the formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof, which is ( R )-7-(2-(2-fluoro-5-((4-(2-isopropyl-2-thiazole-4) -carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzene And [d] thiazole-2(3H)-one fumarate. A compound of the formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof, which is ( R )-7-(2-(2-fluoro-5-((4-(2-isopropyl-2-thiazole-4) -carbonyl)-1-oxa-4,9-diazaspiro[5.5]undecane-9-yl)methyl)phenethylamino)-1-hydroxyethyl)-4-hydroxybenzene And [d] thiazole-2(3H)-one. A process for the preparation of a compound of the formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof, which comprises: (a) when L ¹ represents hydrogen, a compound of the formula (II) or a salt thereof Wherein LG ¹ represents a leaving group and L, L ² , L ³ , L ⁴ , R ¹ , m, n, V, W, X, Y and Z are as defined in formula (I), and a compound of formula (III) or Its salt Wherein Ar is as defined in formula (I), and P ¹ is hydrogen or a protecting group; reacting in the presence of a base, followed by removal of a protecting group; or (b) when L ¹ represents hydrogen, the compound of formula (IV) or Its salt Wherein L, L ² , L ³ , L ⁴ , R ¹ , m, n, V, W, X, Y and Z are as defined in formula (I), and a compound of formula (III) or a salt thereof is present in the reducing agent Lower reaction; or (c) a compound of formula (V) or a salt thereof Wherein L, L ¹ , L ² , L ³ , L ⁴ , R ¹ , m, n, V, W, X, Y and Z are as defined in formula (I), and P ³ represents hydrogen or an activating group; a compound of the formula (VI) or a salt thereof Wherein Ar is as defined in formula (I), LG ² represents a leaving group and P ¹ is as defined in formula (III); reacted in the presence of a base, followed by removal of a protecting group; or with a compound of formula (VII) or suitable Salt Wherein Ar is as defined in formula (I); reacted in the presence of a base, followed by removal of a protecting group; or with a compound of formula (VIII) or a suitable salt thereof Wherein Ar is as defined in formula (I), LG ² represents a leaving group; reacted in the presence of a base, followed by reduction of the ketone, followed by removal of the protecting group; or (d) when L ³ and L ⁴ each represent hydrogen, a compound of formula (IX) or a suitable salt thereof Wherein Ar, L, L ¹ and L ² are as defined in formula (I), P ¹ is as defined in formula (III), P ³ represents a protecting group; and a compound of formula (X) or a suitable salt thereof Wherein R ¹ , m, n, V, W, X, Y and Z are as defined in formula (I); reacted in the presence of a reducing agent, followed by removal of a protecting group; or (e) in L ³ and L ⁴ A compound of the formula (XI) or a salt thereof when one or both represent hydrogen Wherein Ar, L, L ¹ and L ² are as defined in formula (I), P ¹ is as defined in formula (III), P ³ represents a protecting group, LG ³ represents a leaving group; and a compound of formula (X) or The salt is reacted in the presence of a base, followed by removal of the protecting group; or (f) when each of L ¹ and L ² represents hydrogen, the compound of formula (XII) or its salt is rendered Wherein Ar, L, L ³ , L ⁴ , m, n, V, W, X, Y and Z are as defined in formula (I), and P ¹ is as defined in formula (III), and P ⁴ is nitrogen a protecting group; reacting with a reducing agent, then removing the protecting group, and reacting with a compound of the formula (XIII) or a salt thereof Wherein R ¹ and Y are as defined in formula (I), and LG ⁴ represents a hydroxyl group or a leaving group, or a salt thereof, followed by removal of a protecting group; optionally, (a), (b), (c), (d) And (e) or (f) are followed by one or more of the following steps: converting the resulting compound to another compound of the invention to form a pharmaceutically acceptable salt of the compound. A pharmaceutical composition comprising a compound of formula (I) according to any one of claims 1 to 6 or a pharmaceutically acceptable salt and a pharmaceutically acceptable adjuvant, diluent or carrier. A method of preparing a pharmaceutical composition according to claim 8 which comprises mixing a compound of formula (I) according to any one of claims 1 to 6 with a pharmaceutically acceptable adjuvant, diluent or carrier. A compound of formula (I), or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 6, which is for use in therapy. A use of a compound of formula (I) according to any one of claims 1 to 6 for the manufacture of a medicament for the treatment of a respiratory disease or condition. Use of a compound of formula (I) according to any one of claims 1 to 6 for the manufacture of a medicament for the treatment of chronic obstructive pulmonary disease (COPD). A pharmaceutical product comprising a first active ingredient of a compound of formula (I), or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 6, and a second active ingredient for the treatment of an inflammatory disease. The pharmaceutical product of claim 13, wherein the second active ingredient is selected from the group consisting of: a PDE4 inhibitor; a modulator of chemokine receptor function; an inhibitor of p38 kinase function; a glucocorticoid; and a non-steroid sugar Corticosteroid receptor antagonist.